AU2015201676B2 - Anti-C5a antibodies and methods for using the antibodies - Google Patents

Abstract

The present disclosure relates to, inter alia, antibodies, or antigen-binding fragments thereof, that bind to C5a and to use of the antibodies in methods for treating or preventing complement associated disorders such as, but not limited to, atypical hemolytic uremic syndrome, age related macular degeneration, rheumatoid arthritis, sepsis, severe burn, antiphospho lipid syndrome, asthma, lupus nephritis, Goodpasture's syndrome, and chronic obstructive pulmonary disease.

Description

ANTI-C5A ANTIBODIES AND METHODS FOR USING THE ANTIBODIES

Cross-Reference to Related Applications

This application claims priority to and the benefit of U.S. provisional patent application serial nos.: 611330,260, filed on April 30, 2010, and 61/471,465, filed on April 4, 2011, the disclosures of which are incorporated herein by reference in its entirety.

The present application is a divisional application of Australian Application No. 2011245117, which is incorporated in its entirety herein by reference.

Technical Field

The field of the invention is medicine, immunology, molecular biology, and protein chemistry.

Background

The complement system acts in conjunction with other immunological systems of the body to defend against intrusion of cellular and viral pathogens. There are at least 25 complement proteins, which are found as a complex collection of plasma proteins and membrane cofactors. The plasma proteins make up about 10% of the globulins in vertebrate serum. Complement components achieve their immune defensive functions by interacting in a series of intricate but precise enzymatic cleavage and membrane binding events. The resulting complement cascade leads to the production of products with opsonic, imrnunoregulatory, and lytic functions. A concise su mmary of the biologic activities associated with complement activation is provided, for example, in The Merck Manual, 16th Edition.

The complement cascade progresses via the classical pathway, the alternative pathway, or the lectin pathway. These pathways share many components, and while they differ in their initial steps, they converge and share the same "terminal complement" components (C5 through C9) responsible for the activation and destruction of target cells.

The classical pathway (CP) is typically initiated by antibody recognition of, and binding to, an antigenic site on a target cell. The alternative pathway (AP) can be antibody independent, and can be initiated by certain molecules on pathogen smfaces. Additionally, the lectin pathway is typically initiated with binding of mannose- binding lectin (MBL) to high mannose substrates. These pathways converge at the point where complement component C3 is cleaved by an active protease to yield C3a and C3b. Odu.-r ---:)dn*, ays aet-vuimo complement attack can act later in b-e sequence of events les-uia-c, to rations aspects of e-.nipleroeni funeuoit. C3-a ls ssr:ds5apfS5?Mebrn. -and- bthefceiis,: M-wèli stWlé certain viruses a-ui hummie centple:<es, and uses them lor remo\ al lfom the circulation. Ob in tfïi-' role 6- known ns opsotfut.) The opsonic junction of t'ëfc is generally costs klered to be she noes important ami-ittieetke action oi the complement system Patients wish genetic lesions that block (Co function ore prosie so infoe’tion by a broad em-set} of pathogenic organics-s, while paticsiM ftuth lesbons laser in the cotnplement cascaelc seepienec, i o , patients w-i-h Icsiona nun block C5 functions. am found to Bé ruose prone osily soil then only somewhatmore prone. C3b: also: losing a complex with other components tmlepte i# each; pathway to hosss classical or alternative C.S core,·erusee.. which cleaves tbs inn..- C5a ami C3b. i. 3 C thus regarded as the central protein in the eosepk-menS reactie-- sequence sittce it is -êsséftiiM to both: the pathway $, dins property of CI3B i$ regubsed by the sensm profeam Facies k 'which acts on Ob io produce iC5b. While still Ihoctipnai: ass opsdnio, l€3h #atkMtMh:-M<jMV&amp;€5 eoiieerisse, C3 is a 190 uDa beta globulin found hi normal serum at a concentres-on of appro imatek "5 uc mi sO.-i uM). C5 is glycosylated, with about 1.5 to 3 percent of ltd ttsaas: atMBhteddO: caiWohydram. Mature €5 is a hclerodimcr of a 999 amino acid ! 13 kDa alpha drain shat is dtsnMde linked to a (-33 amino acid '-’3 kDa beta chain. C5 is ry tttbest"od as single chain prcesovmr protest! product of a single-copy gene :fflamiihnd:et ah. 119911 Jlmmmèt 14?».;362-36Hl Tito cDh.\ sequence oi lltc nans-mapt ol this gene poo he--. .; seem tad p-rc;-C '5 precursor of 1665 amino acids alone wssh ast id snttno void kade-' sequence ise<\ e g., L .S Patent No. 0 555.345 r 1 he pro C'5 preeurset ;s clear ed after amino voids 655 and r>5t;. to y -old the beta chain as css asnme· terminal fragment imn-m.· acid residues + i m 655 of she above sequences -md the alpha chain us a i-urbosyl terminal fragment s amino acid tesidtses (-O t-! ) (Ns of the above sequences. with tour antl;n.> acids t a on no acid residues ό56-;-po ot the above vcquencei deleted bewvecn the tww F5u is cleaned from the alpha, chain of 05 by cither alternative or classical C5 eoitvcrtaac aa an atnino tenninai fragnient cotttprising the llrst 94 anuno aci-Js oi iitc alpha chain O.e., ;.unu'u.> acid resis.htes i:>60-''55 of the abov-..· scrpo'cnc-.o ,-\pproxitnateiy 30 percent of the i I kkbt soav- of C5:t is ^m ibusosi t-.- csirboitsdraie. 1ï!ie öl$«v&amp;ge:si;té «Cor immediately adjacent tu, amino acid •^iiuè733-ófite'a&amp;vè'-^tlê{3èé.. A ééïripMiê thw wmtfd bind as, ur adjacent, ίο thlc- cleavage siic would have tb,· poic««tril to Nock access of the CS eonvertase enzymes to the cleavage sue noi thetehv >-.ei a\ a (.ompienum inhibit-e. CÖ can uKo bo eem a red d>, moans othet than C5 e- me eoase aem tn i s mi tod tryptdn digestion (see, c e , Minin and Man ί I 'ft M J ƒ?<?-»; v>?· >} i 1 ;b i 5if *- 1 602 and \Vetsei nod Kolb iokC s./ j[2s'?.?(bf--.?.?SM. thtombm, a; ui mud non on cm (Yamamoto and Co mure t / -.?<?.->; \ "O pup;. and ilamerm et a! ·, fossp

Mui<'C ImimuKii 26' ϊ I on 11 -12 ° can d--'· CY'a\ C5 and produce note, c (Mb f leavago ofC5 te'c-iSe··. -' êa. η η<man m-apfo Into on and chansotacSk factor, and Cab sCsieh th rough a sot to · el rna -teut innamen on a lead- to tits tormanen of the ivnc termbtni complement Complex r 'foas C ha and ( Ced g(so ba' c pletorropst ·,nil iKdYhiiBg.pröprticy, ^ :ti^i|fy^g.0ig".3:etesc of dorvosiressh isjflammstory factory such as hydrolytic erury must, reactive oxygctt^^Ies^.!a3t^M#ö:hi0^chi metabolises and various Cy t--kmcs C;fo -.onibmes o uh Civ Co and CS So form She Ob-S coniolca at the mrCee t.-f the target odi. Upon binding of several CO molecule,*, the membrane snack complex t MAC, Crth-9, terminal complement cotnpimv-TCC) is iorated, When sufficient nurnbera of MACs inacn into tar,act cell iricmbmnea the ojxming« they create ; MAC pores) mediate rapid osrnotte iysts of the target cells, fearer, stott dytlc concentrations of MAC?· cars produce other effects. 1st yarn en I an membrane insertion e-ί small numbers:· of the C 6b-v complexes into η ruled red nl cdla and j-dateleta nan cten-e deleterious edi activation. Iti mane crocs ado alien may precede cell lysis.

As mentioned above. Chut and C6a am anaphyIntoxim.. These aetenited complement components can bigger mast cdl degnmsiiabem which rcleaaca histamine from basophils astd trtast colls, and other media-ora ef inflammation, resulting it; smooth ntuade cottfraeiion, incivnaed \ascuiai permeability. leukocyte activation, and other inOarittrtatory phenomena including cellular proliferation fesuhmy in Itypereedulataiy C5a also functions as a chesnon-sene pcpt-dc that -mm os ti.i altraef pro-inilainmatory graanthacyica to the -die Ol complement activation. that: receptors am found ost the surfaces oi bronchial and dveoku epbltdid edlb and fjmnehial smooth musel# cells, C6s. receptpiS have also been found on cosit't-..>phds. tufisi cells, nt-tne-eytes, nentrophils. and activated lymphocytes

Any discussion of the prior art throughout the specification should in no way be considered as an admission that such prior art is widely known or forms part of common general knowledge in the field.

Summary

According to a first aspect, the present invention provides an isolated antibody, or antigen-binding fragment thereof, that binds to a free C5a polypeptide, comprising a light chain CDR1 comprising the amino acid sequence depicted in SEQ ID NO:20, a light chain CDR2 comprising the amino acid sequence depicted in SEQ ID NO:21, a light chain CDR3 comprising the amino acid sequence depicted in SEQ ID NO:22, a heavy chain CDR1 comprising the amino acid sequence depicted in SEQ ID NO:28, a heavy chain CDR2 comprising the amino acid sequence depicted in SEQ ID NO:46, and a heavy chain CDR3 comprising the amino acid sequence depicted in SEQ ID NO:47.

According to a second aspect, the present invention provides an isolated antibody, or antigen-binding fragment thereof, that binds to a free C5a polypeptide, comprising a light chain polypeptide comprising the amino acid sequence depicted in SEQ ID NO:42 and a heavy chain polypeptide comprising the amino acid sequence depicted in SEQ ID NO:45.

According to a third aspect, the present invention provides an isolated antibody, or antigen-binding fragment thereof, that crossblocks the binding of the antibody, or antigenbinding fragment thereof, of the invention to C5a.

According to a fourth aspect, the present invention provides a pharmaceutical composition comprising the antibody, or antigen-binding fragment thereof, of the invention and a pharmaceutically-acceptable carrier.

According to a fifth aspect, the present invention provides a method of treating a human afflicted with a C5a-associated complement disorder, said method comprising administering to the human the antibody, or antigen-binding fragment thereof, of the invention.

According to a sixth aspect, the present invention provides use of an antibody, or antigen-binding fragment thereof, of the invention in the manufacture of a medicament for the treatment of a C5a-associated complement disorder.

Unless the context clearly requires otherwise, throughout the description and the claims, the words “comprise”, “comprising”, and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of “including, but not limited to”.

The present disclosure relates to, inter alia, the generation by the inventors of a series of humanized monoclonal antibodies that specifically bind to free C5a protein (that is, C5a that has been proteolytically cleaved from C5 protein), but not to paralog protein fragments free C4a or free C3a [the antibodies are, often, referred to herein as anti-C5a antibodies or anti-C5a neoepitope antibodies]. As described herein and exemplified in the working examples, the generated anti-C5a antibodies exhibit a high affinity for free C5a. For example, all of the humanized anti-C5a antibodies described herein bind to free C5a with a KD that is less than 1.25 nanomolar. Many of the antibodies bind to free C5a (e.g., free human C5a) with a KD that is less than 300 picomolar; several of the antibodies bind to free C5a with a KD that is less than 100 picomolar. In addition, the humanized anti-C5a antibodies described herein also inhibit C5a-mediated signaling. Further structural and functional properties of the antibodies described herein are elaborated on below and exemplified in the working examples.

The inventors have also demonstrated, using an animal model of rheumatoid arthritis (RA) and a surrogate anti-mouse C5a antibody with properties similar to the humanized antibody counterparts, efficacy of anti-C5a antibodies in treating RA. Also shown in the working examples are experiments demonstrating the positive therapeutic effects of a humanized anti-C5a antibody in an animal model of human C5a-induced neutropenia.

Accordingly, the inventors believe that the anti-C5a antibodies, or antigen-binding fragments thereof, described herein are useful in a host of diagnostic and therapeutic methods related to disorders in which C5a-mediated signaling contributes to pathogenesis. For example, the inventors assert that the humanized anti-C5a antibodies described herein are useful for treating or preventing RA and other complement-associated disorders including, but not limited to: atypical hemolytic uremic syndrome (aFlUS), age-related macular degeneration (AMD), sepsis, burn (e.g., severe burn), antiphospholipid syndrome (APS), acute respiratory distress syndrome (ARDS), inflammation-related pain, asthma, lupus nephritis, intrauterine growth restriction (IUGR), HELLP syndrome (Hemolytic anemia, Elevated Ever enzymes and Low Platelet count), Goodpasture’s syndrome, and chronic obstructive pulmonary disease (COPD). Additional disorders that are particularly amenable to - frenmmm with a husnu-nced smh-Oéa amibodv, or ssmigcn--bmding fragment thereof, are f stoxn m the arr and reenedhefUm. the humaubed asm-Oha amibodte,.' described heren- feature a number of <kb >mu..!cs. e ··;, e-\u‘ terns, dun bind to -isid inhibit elesnage ol. suis-iemuh of mimste t I ;ko amh ngem.·, the anu-t. 5a antibodies pand '.i-mg-.m-mn-.lmg tr:t§me$ts thetv'ffi do,'cube*: heresn jk· capable of inhibiting the imaphylaiONse doxrknvtun effects >u t 5 ,-χηχ >u;on as mediated through <d> hsigmest? C>;·!. fhat h, the ambCSR am shod sc,·· describees hex-n < a» inhibit the Oha-med-med -rdlamm-afory response, xiueh is \u>-x : -o pi-λ tü infegrai part sn the pathogenesis of eompk-mcm associated disotsiers 0u< b n->, but inn (busted to, sepsis, RA. and asthma. However, as the cosne-P rat-on oft. > m hen ;,n· as. rum :s approsumsueiy Ox ? u.'vl: Rax cl and ibmpburn ; Odd i i./ /u’xtxxs j oo. 4 s. do A-the »sc oi Inch concentrations χ; j os frequent administration on and-Γη antibodies :s oin.n necessa-v κ- eifxnh aU mh-hu (h'h, and thereby stimbu Mk C>>ms mated uifiamniatorx tetpesv-e. sn a Sin-nan. Lxdd· e Oa. On is present tu bi-; »d 'tt s rased ixxcr e- -neenoritiosis -md n> oben re'-toeied to specific areas οΠοαο contrLuixru nose. ash-si suets as, e e,. the lungs m sstimu paai cans site joutt,' of R-\ panent', -η' the -knx.a »u the eves < d p-ittents xith AMD finis. -he antt-Oa amheodic.·. dose tabes: hen. -n can be admbixtxed < e a . locally udnisnn-Jead to 'ties sd complement .klxxsdrak t<- a human at a touch Soxcr J-nc anti or tess iVenncnti) than, c g . at- atiu-P '· .nusbod’-· and offecthch pros ids' the same or greater inhibition oi'CÓa m a hus-wo, 1 h<. ahthu to adnuraster a lower desa os'the cun (.’pa tititto'OC. as ctnnj'ared t i;· tee muuued ο οχ a -; ., ;-o o ? x y a t txi t a a . ra s du-avs a .a adduionnl denvesy >-rote.' ent.b as. e y subeutanC'tns adi-entMrntsurh btintinnaetsltn' ndntniisjtntiojt, isurapuinuanny doit',ere, ami nbmhosnaikm \ i:. tue -:s·, vfbi,dopii..adx skpradttbte nnct'-apheres Λ Soxet coneeniratf-it oi annaett ( '<u χ et a ns c'5 also Pt v os s a loupes balf-dt'a a-h tit a tun i-if 5,; ,nu»bod> as compared to, e χ, the halidtie of adèerapenttc atniibpdy thatiwgeis terniina! hPtTtplempBb due to a reduced eónffÉtitbbti oi amicoa -ntCiittncd anri-i-a-dy c‘care tree hi S:ddi:i:.:u, the atifi-Cêu muibodtCs dcses'ibcd heroin cun alao be dtsttngusshcd isom thetiipeette agerUs shat usitahtt terminal complement (such as <f5 irthibisoisi by tb.elr sal civ protue. Λ uotsbic cottscni utticc nsi" inhibiting terminus compl-cmem con-p'Snems such an (da, C5b, ('h, C7. CK. or Cs1 is ticcrcascd prontcnot; by she lu>'-t inumnse sveiasr: ssoautst the cricap^uhusad bncieria that tcrmimil complement orditutf ily lyses lor söd isiefcsemi :go:nètmomi&amp; $%β,

Baessey ei al i fMöt dih.&amp;tpfmmmbl· 40:1 OSM and Btoslsky (200ό > Bmod i 13(26):0533-6527 As tbc assn-CSa antibodies inhibit the i' éa-mediatcci i;sliummat:.>rv response. fcm do firn prevent she for; nation of the terminal coinpiemenf o o moles dan lyses sixsse oneapsiuased baeseria. ps.u terns ssxxnVmg a therapeutic ami·· C5« ami body described bereis; ox .-uld nos requite a profeeth e vueci star son. eg,, a vgoeibalios: against iiii&amp;MëMsêiïé· gMwriuimië,- .Abèiafdibgiy; in dit© sypobf tie et amipen-binding iVagrnesu thereof Hun hinds so free ('5 a. in vsroe embodiments! tbc antibody :.-r ardigciphindina fragment dsueof binds U; ilee human CSa (hC5a: e.g„ a hisnssm CSa ponen; eompriaisng or consisftsig of the asninu aeid soutsutjce depieted in SFQ iD NO:i ï. in ''urne Ci'nboduïsens.·;, she antibody ran b:nd so a dosas'gimuod form of free C5a, e,g., the desasgirsaxd fonts oflnsman C5a comp-rAing. or consisting of, the amino aeU sequence depicted ns SFQ iD A0:2. The antibody can hind 1o a ngbepitopd of' if eg OS a, w hielt: epitope jsr stbt présent on. bbdlèavèë OS oils pfesbnt όη phly a mlttor tïaetiöit of tot al anylep'gdl O 5,

While the dwelvstsse ie in no v-ay limited to assy partsesjla? thorny os moebumssn ui'net km. in -.ome esnbodnssessts, the smsi-f'Sa antibody or mnlge;·-binding Swgmetif tberdef birkilr 't® fteè OSa fe;g.,, Jibe %GSa) and may als&amp; histO to a subpopsshttion -2'osxheas ed psoecssed OS pr.g,. plassssa OS; CuuoUuhug km lisas; 10 (o.g.. lem thisss 0 S. '1 a S, 5. 7.5, ?, oW 6. SO, S, 4 S, 4, OS. .1 .).5,.?., i.'l j, 0.5, 0.4, 0,3 i 0.¾ or less fbass: 0:.1. 111 of ibg tplilpopulaiioo èf lull loigtlt Qf IP a satnple |ó. g.y a bk>od :;r plasma sample os a -.ample eontprking reeombinusd full iengpis 05), n lsich -nbpopuhdion 1:- in u hok or sss pan. denasntvd -usds fan ass otherwise oodudod OSa svoopstope to e-hich tite smi-Cca antibody or dunce;n binds. ;s exposed Thus an riUit' f.; antibody or amigess bissdissg '(ragmens thereof des-enbed herein enu. -t; aonse eoibodisssenn;, bind so Pee a Sss, bs.it stot x> bse uneiessred ('5 protests of the f’O'f. or gs’eater ussdeased, suative 05 p-spstUstsots. in aoisar unbodimems, she abio e-deaeriben past ini h or fully denatured yubpnpukfion efs'S ia sssaeslxc <>·: lux redus;ed aetivisy te.g. leva ihsns 90. SS, Wh ~'ή· "0 b<i. ta). 55, 50, 45, 40. 15. hg 25, 20 15. 10. 5'x of She '-Kiix igv t>( bfly-f;aseiionaf fsiii -iessgth 05 piOtessss sss assy number edOsssufok. aasap nsefti! for testing· €5 activify, g,g.,, a hesnolytie aassy of adISÖeq essay!: (see below). ' tttbit%ö#l· for the aefivity of the mister sttbpopdlatlpsi: to wbieft &amp;n &amp;nU-05a antibody described herein may, in some embodiments, butd are known in the art sand desèfthed herein. in some embodiments. any of die anti-CSa antibodies nr mdlgcmblsiding fragments ftbf inhibit CS agdvity Ins«ft-ffftp^6$tóly$sr· assay or an .¾ Woo CHöOoq away exen m -be presence of at leas: ^qual to, or greater titan a 5 <e.g., 5.e. b. 7, ,s. S', 10. 15, 20, 25. 30. 35; 40.45. 50. 55. Hi. 65. 70. 76. SO, h.\ u0. 65. iOO. Π0, )70, 130. NO, 150, 160, Γ-0. I HO. WO. os 300} -f«4d cxcc* of the anti-Cfa annbody or antigen-bindmy fragment thereof to nndeaved ('5 (,e.g.. uuekrwcd, native C5; In some smb<‘dimenSs, any ofh'he ami -C5u antibodies ot mKigen-bhiding insgt neats thereof described herein si is not inhibit C5 activity in an /.<? v r\< ίιοηκ'Λί"}' assay er m t tOuasae. exec οΚΌ'^,,ίι'Ο^ΗΛ'η ah< -ui a 5 - ibid io 200- ink; sc.g., between about 5- fold and lOO-toid. between about 1Ö-föld and ICIisMd, between: abodt :20-¾¾ b.nd: ifesut 1 Of fold and 150-Add) excess m the asui-O 6a antibody or antigen -binding fragment thereof to uncleaved, native Oe. Inhibition, e.g.. as it pertains to 05 activity, nscindes ;n least w 5 0.:.0.. at least a o. W K, 10, 15. 30, 25. 30, 55, 40, 45, 50, 55, or 60- % decrease in the activity ofunekuwod. native 06 ay e.g . a hemolytic assay orOHOOeg assay as competed to tlte etrcct -,.>ί a control antibody ior ;rm; gem-binding fragment thereof} ander similar edn dit l oss and at an ègmmoldr concentration, Stitetasrttal Irflbbiiib», m used herctn. tsters to inhibition of a given activity (e.g., ofC5 activity) of at least 4t) gè,g., al·lease45, 51),, 55,61),:65,:3(1, ;75, Μ, B5, 90,-or 95 or greater) %, In some embodiments, the (‘ s is obtained from plasma, (e.g., purified from or present, in plasma, e.g,harnaa f Idsma),

In emoe embodiments, the antibody or anttgen-hmdhsg fragment dteseof binds to: a 05a protein fe,g., a batnsst OSa protein) with a Kp that is less than. 2 nM. 1» some embodiments, the antibody or antigen-binding fragment thereof binds to a CSn protein With a K'o dtat is less than i nM [aUo referred to herein as wmbrnmomoiar aiiiniiy'b. in some embodiments, the antt-Cöis antibody or antigen-binding fsagmesti thereof binds to free C5a with a sebmmomo.iar a filmt y [e g., a Ki> of less than nr equal to 9.0 s 10 i! le.g.. less than or eqgdilo-f x, J0 *", l a 10 *‘3 7 χ ΙίΓ^1, ?S x iff !0 5 χ 10 m. 4 X Iir'* 3 x iOvUs.2.S:x 11318,2x ΙΙΓ1* 1 x 10 5.0 x Iff41, X§x KWh 6.0 >. 10 u, .5.:0 x. iff* 5 4:0 x !(F M, or 3,0 x fff * *) M| in: the pmsenco of a molar excess of anoiesved, native 05 f e, g,., purified sadfbr reearobinaiit CSx In some eiBbodltóetits, any of the ann-05n antibodies or antigen-bitiji-tg IVagmenis themef described herein ha\e«t kast « NO-eg. a* learn ΠΟ, 120, KM», NO, NO, 160, 170, 100. 100 "100,

:0:5, :50. r\ 500.4«hi, 500. poo, "on, ^oo, 000, 1000. :000, smi 4000,5000. #0CX1 lèÓ^. 'Skater »ll»ily.4-ei'g;, -represented by its K.N for bve Γ kt dons Nr eec kneed, siaisv..- ( .5 protcsst.

Nos, m another expect. Ok disclosure feature.'' an antibody or antigen-binding kaguKK these N'that (to bln# to free ( 5a ;c g., hC5ai w hh a ruhstanooiokn affinity und - hi binds to free C5;: n bh ;m stfikN lb at ss at least 100 (e.g.. at leant S10, 120, NO, 1 10. NO. IN), NO, ld(i. 1 VO 700,.' 15, :5(S, .-75.300. :00,500, Nib, 700. #)0.

Vi Hi, 1000. 2000, ,0)00, 400(5. 5000, 0000, Nil# dOf>0 #00. ov 10000) -fold greater lhan Os ο- mespotnUstg a ion it)· lea unckaved. tsainc ( 5 protein. for example, art atilt-0 >a antibody nr rmlr-ee-Nob;eg fragsiann therool dosesslvd herein cast, in x;ine e-ntbediioettis, bind u> free bC5a w nh a K.. <;<f 1(50 rtM and to at leant a ssibpopubttsurs of unckcned Inanan <,N protein o lilt a K,.·, -led is ai leant lOO-foUl higher t'e.g.,, at tests! !0 itM)

In &amp;not ν'.ι,'ρ,νο ov Jtsek'sov tearures m sv dated tons bo# »> ,et'to,> n btndlrtg: i&amp;agmbttr tberebf fbai bio# to a free IrosMsi (25¾ tiplyiieptile ftavingihe smtino acid stXiSKtKs «Lp:c*ed 1st Sf '2 ID 'Ob, wjvtem Ok atsi#ed\ m ustngeo binding fragment Γκο,οΐ bines to tbe om-mr t #. jvb peptide nidi a #, dsat ts les' ibnn ï .25 a 0'*' M ok o'c-e'Vc ot a molar s/sec# e,g , .¾ 2, 5. 4, 5, ·,\ ", 5 -K 10, IS. 20. 25. ># '2, AD, #. #, "o, SO no, too, 550, 2oo. ?0t\ UN ot 500-teKl stsohn esee.'s: aNncNe, oe, ere no kuna» t 5 - r, •-r hum,ui i N ; ## p, et wire stvibodimcmn ; κ nusi'·2,\ et atotgvtt lundutj» it '.mnem Oicto *'bsts# me bee nk5n. polypeptide wbh u .'UNunentcks nUN# se e , ms\ of the subst,mmjvk; -v # teenad hesnitt) tit lire me.-evee ot a·, least -a· greater th n;, a 2-ibkl nndai e\e<'-v., mf !}o greater or less-than a'OP λeOO, NO, 40<>. «o sop ;a{) u;o,->0 St)., 70. 60, 50, 40, '5s N 20 o' N'> hrsoln .'λϊλα of unelese,ee. bee h('5ii. In smite cnibiKenente, dte asuibody ι,-r atHigeii-btnduig bagraerd rhetcol bnttb to a free ICSa poiv|aepöde a snbhwh»^!# affinity (ο#>< any of the subnonortKlar kp'a reeneb hnrentt in the ere settee n.f benveen 2-fold astd 20-bed rnennr escess η:·ΓuncScmco. r-Kive ('5 eser bee h<25a. lit sonic icrnhodimettK, an antibody or nubgcn-bcnbng iVagsnciu thereof binds to a bee hC5a polypeptide with a §«batóömb)iar.alffi6%'(!öjgg.^y oftbe sbbtiasipmplaf Rtt H fecited Stereiti:) in the presence of between 10-i'old and 20-fold rrmiar eeeesS of ns te leaved, nalive C5 over free Ii05a. In se-mc emb-ktnients, an aïtiibody or atttigen-bindtng fiagment tliercerf hinds t>) a free Md 5 a nbypepn-f- with :- Mdm.uvniXihir .JV;nny ie e., too eftM.

Mii« :;mom<<br k;Λ iCGIi'.'d h'-'i'OtO) Kt ïl'sC presence <jt' bcH\ CO!! 5 fold fïï J ί c- ibid mobs exec·-»;·· of nod cm cd. mod e ('3 exu fme hi.'5a so so sue em^amnusts, an amt bods or -nkgembndnn;:; fragment mmved' husds ίο a in.-e M 5a poh nephdo wish r subncsnomei tr .dt'msf) p,- g am o' the s-:bnanoondar Ki.\ mestpd herons Hi the pivssnci, -a as kaxt 2-fold, hot ;<· v<;:v; than a 20-Odd mok» exec--,' ei'smdemcd, n.n e,».' e'5 os ar free I o '5a Sat. h men-am me sns ca o bo in » /? a' nteas»: esssenl·· m.rne. >. μ s- sti-iard mfmity dek'tniinanon ics Ιηηρικχ·', mans o-'tshieh arc re·. w\S and m described herein. in another»\peek the dbdoxnre features· ass bdaied astobody or mud-ess-binding fragment shewn! that bunts So a human ('5a pnlvpepode having the ammo is>vid t-otp)etscc Uepkieu in SbO if’) \:0; t. wherein fisc anbbodv or medgen-bitsding I mewses η thetcs.-f binds w the human C5o polypeptide with a bd hun is lex- than 1.25 is K) '' si and w herein the antibody or asswurndvadaq:-. imgment thereof does not subsum! tally inhibit, «x consparest to an eqahrsohu' amount of ti eoasswi aniibody os-antigen bindn;o. fuapwea; thereof, C5 activity even in the prexesioc otdk'W bum. or equal to, a JO-fold ns-da? excess oi the asodCdn antibody ot asssseen-busudsg iragsaessi thereof so vase leased, stadse d.;d hi some embodiments el any J'the anti-C5a antibodies or antigen-binding hugments thereof described hewus, the antibody or assftgmuhinbing fragment tiscrevf hinds t- s be·:.- human fdu and i:- cross-reamhe with free 6'Ws front at Seaxt one tson-Innnan morn no bon species I'et evnmpk·. its -wre embodim-wux, att ant: -Cda am foods o.-r sauipua-bhebne fragmesu ilkreofi bits Jo to free <f5a from huts tan se μ., svitfs sSïbnattojrudar ..tffinitys stuf «Ko binds η» tree t'5;> frt'rn a ο-m-nmnno primate fe;g. s egö-ömsaigtty mscaquds stesgfr ttsscaqpg,. ape. babooit;, cbitsipaiseee. orsngstatt. or gerilfiO, a rodent te.g . moose, nn hnos\t<rr. Ouinen pip, or rabbit), cost. goat, donkey, pig, dog, cat. or doom. In some embodiment', an moled5a antibody or runigen· binding fragment thereof described herein binds, to itee hC5a with a K.1; ofleen thou of equal to 1,9 xiJO'tesd ihaa or eqnt.tf to 9 x :10’:!χ. dx s.f)':::. 7 x >5 y (0 ! 5 X 1(S ud 4 i iÖ'i;o3:K i:0tSii 2.5 X 10 ii;, 2 a SO : f S x HV^ 6.0 x 10:'! d.5,0 y Hf:d or 3-0 χ ΙΟ:ΐ5ϊ) M and tp--lïee ¢5¾.¾¾¾ eystotyio!ttn,s trsacaque (or another non-htsman ps'insbe spes'ie.s) v. hercits the affinuy te.g., represcniicd b> its hm) lor humass C5:r &amp; no. more shim 500 te.g , m- tsa>re that» 5. it). 20. 30. Js; 50. kO, 70, Sty 90. 100; 125, 150, 200. 250. 300, 550. 400. 450, ur 475) -Γ= ?kl greater than ila.· a Stab's for cynomolgts\ macaque ter other mur-hummi primate s-wesoa; C5w For example, in amne embeds meats. the nets 05a antibody binds te Fee hv’5a with an auirmy 4 tat w ret ns-re '-hasi 50-told greater that· Use eop-espimding «tunny el the amihe.dy so; aetehurwm pritmhe 05a : ew,, a K;. for free hi” ê<a eh Hh* idvl ;.auj -, Κι, fur mm-hwTtw; primate (55 a ui'ne· roe-re dm a 5 nM5 In some etrsbodhnenK a;· aarwdha anttbedy nr ssdwasehinbwa, Fagntctu thereof described heroni blndr to fro;. h05a v-irh r K» oi'tess duro or equal So \ 10 ,l ,'e.g . leas than of equal to § x !10:i 7 &amp; x :ffi':R!, 7 X ÏWwy-$ x- 3 Öï0. S k Iff *7 4 x. 1:(07. $ x IC0'7o.5 ·% 10 0 2^ 10 0 1 V. u>H0\ !{ί!\".0Ν 10 0 6.0 < ffi 0 5.0 s. 10 0 4.0 \ 10 ; dor 3 0 \ 0) ') M and also hmd>· to (' 'u Stem a sodessi se.g., ntmme, rad nr mhbstq oiterehr slse affinity te.g . repièaensed by its K.r lor itsm-m; C5a 1- no mere dnm 1000 Ova,, r;e more dnv 6 01, 30, 30, -Ki, 50, ;Ό, 70, xs\ 00, i 00, I 35. 1 k), 200, 350, 0)0. 550. 400>, 430, 4~·} 500 550, m;0, 650 700, 750. Si 10», 550. ·χ*0, li50. ot o“51 -ffiid grtuster dra a the offdhi·. Oir rodtrsn i 'ba in some era kali meao., aas ef dm a ?uaf kt anbb-OÓièi or women -bm-htna freu;moois dvmof'dv.erhm-d harem hdad s\üb aahromiamaha adiaay to bods human <55a andfeC'Óa from a π-m-human mamma! <em.. 'i p\km or a nuu-hurmm prims te such as cynomolgus tmteaque), An amihody er aabyon-bindk-g fraymem thereof ear). in some embodiments, baai to hornaa C5a and nou-hunum primate C5a wish equal affhhn ie.g.. an eet ui va tem Kod

Bor Μ*Ητ$$· an antibody or asffigen “binding fragment: shereoi dra: binds re free ha war a C5a wish ^ubnanem-dar affinity fe.g . a Kn of los' ;fbsn Of ogKiO; to Β,-O s less: iimrs brlequal la f x:i0'0 4 s 1107 ? x 10" '\ O x. i:ö"iö, 5 λ Iff·'·, :4 X 3;,x ..if10, Β,χΐχ. iO'rS:iv2 x ICr^yl X 0.0 x lO^l, :7;Β:χ: 10 ::. 6.0 x 10'!!. 5.0 ,s. 10':0 4,0 x er 3.0 s .30'**1) MJ ansi is e:msa-madóvé:k%h ffee G54 fmm yynomoigrss mac^eibk^her.-tmft^ttfwn' onmaiy}-: the antrbmiy or aiHigcn fandiim imernmr; duneui binding to oynormdgas nnteaqae tor ether nonhuman pritnatc) C5u «dit a K-rmst' less than M) x 10 ’, x 10 S \ 10 7 >- 10", 6 s ery 5 ± 107 4 X 10 .1 x i0'7 2x10'y 1 x 10'h O.Ox H)'S:i scar, tesy thns): v x 10;7. K X 10'^. 7 x 10"·. <! s. iO'!e .5 X If'1\ 4 x 10--73 x 3,5 x 10'0:2 x 10'Mi, 1 x 10' :K2,or:b:,0x.l:fn)::Mi, sObèrein: tire sISmty Ibr: Ita.mi0- CSt -fa tro more itmn: 500 fe-g,, ne mere rhart 5. 10, 20. 30, q{t, 50, t'O, "0. 50. SUi, U'tft, 135.. 150, 200, 250. 100. 550, 4M, 450, of 425) -4014 gf ester that) tite affinity for eystothdips manaqtte for non-httsnan printsde) (35¾ ;e.g , Ole. ter In;man C5a of 100 u.VS and a K), for neu -hamai· prinrsfe C5a s>f n<t more dxm 50 tiM t. Smtable methods for dmerrrmime the aftlniry of an antibody or nmigeti-buxlifiii fragment thereof for a ghen anogen are known in the or? a ltd described mss) oemr'kiled herein. in iome embodiments the eruss-rcaeisve ardv-Ohk antibody or antigen-binding fragment thereof fnoetioiUiily inhibits boh: tree hChu and -he non-human nyamnnman Cha to which if bind', for example, an antibody inhibhs by at least 70 tc g.. ;.e icuaf "5. St\ l\\ oti oy os ^re-be·'! ''·- imutan C5u-dependent human ueetiophil adivaliutf at a ntolar mtlo ol 1:1 tantigen-binding Μοοί.'οη> ami inhibit'·. by at least 7bis.g.. at toast so, 85,0(1. or oy os greases·} uondmroun mamma Iran ί'pa-dependent neuir'-j'hil ;x ovation < d:e neust oplnl? being bont the mme species aa the non human tuanimailan Cos to which she antibody binds': ;u a molar rati·,· ol S: 1 f euhgembinding witetibia!. hi anDhoi osf\\:, the dwef-nne ice tin's-, an isolated antibody <u antigen binding ftagmem bn,:root dr-sl btnds to * :>v.e isCha puKpepJtde has ing Sits annuo acid sOijuetu'v depicted ns Si D {[) \0 g v, heren: the nmtismb of unSigeu-bsudhig sVagm ntdie reel bunk to fee atm-.m t '>; posvpcpnJo v·, uh a k; tint: e> less that· I ’> \ tO ' \1,5 fid v\ ftetem the antibody e .unigen bind nip tragn-mu ihereobbuKls to in bli id ha and se t ha Irons a smii-bumim tmunnkdsuss sivcses The non-hwnisn mammalian 'ffcctes sat- be, e.g.. a η on dm man prinute sot h as m υοηνηρυ- macaque, rite-us macaque oi baK«·» In s'-me enslnniuisenr;, she non human mamma hen species is a sevknl -i:sb in nr aso rat. lubbrh < musea use, perch, ot ham.-tef = u some esisbodliiiesoN. the audhodv o; .mdfg.m-Hodifsg fragment die rees binds so Id 'ha with ast ;dIn'liiy no gs\amr thatt sOO-told higher -ban the emsesponding itftuiify b-r kca Irons the con-human ηηηηηίηϋιΐίΐ speck·». In -"we embodiments, she antihsh <« aungvu-bittdinp fVagmun inhibits b\ ;n lenst sit",, human vhkuuepunuerU human nenfrophii achs ·ηί'.ι;ι af ά i-u-ku' iniie ot 1.1 iamspen-bsuduin die On: in soiiso embodmienn, the antibody or antigen buiding bagm-.-m memo* binds to tree 05» tV-m a in-n-human primate :e g , a e\ nomolpu- ntacacpte - a she'-us macaques the free s’ Ό pteteh: hex mg .ui animo nmd 'epuei^e comprising, or consisting ok site amino arid sequence depicted m Sb\> ID VD i m ki-O ID DTÖrlSÓ, \ίΐ described tit she working examples, she inventor? have also discovered is hiv&amp;k'u? ant 1-(. "5a antibody, BNJhH.k dial binds to tree bo a (in (his ease humim b'l.Sa) wiilkMgh aSImty &amp;pi|, wisbif sktcli fowif affibjty, :nBeibSi.kgd: llustms Ci:51;h€S}* wherein, in a composition sc.μ., an aqueous solution! under physiological conditions fit CsUitlibrlum, und k the presence of., nmlur excess of -we teased human C5 :w L-ompared to the molar arnotun of the .mogen lending xiks ol the anneodicv at leas, 95% of the plurality of mu mod km ouch bind rso mote than one info molecule. the second ankgemhwdnw win -if the at lens, k'R, et 5I10 plurality of antibodies romama avfidabk' K'oJ·· subr emiia'd’·· a·- aaahk'> to bind to free (''ha. Ά hate the dock a nre 5- in no way boustd fay any psrtMular ihepiy <<? mcchautMn ofaeeon, the mvenvrs behe\ e d;;n ike bleak;·; ant-C a··, nnur·ab blok to mscicawd c.'> in such a way (c.g., at sock an op-tooei that ;u,r\' hindrance precludes or at leak aaki.aoiaHy inhibits, the binding of the see bail aatigea-kindtag site of the aait-CiSa hiihhptty to kyecom! urseleavcd 05 protein, although the antibody can easily accommodate the binding to two k€5a moleeuks, Thus:, the antibody, oven in a molar excess: of uncleared :05, retain;; the ability to bind to free 05¾ -a kh high affinity and thereby retains, even in that molar excess, the abbey to inhibit the pro- inflammatory activity of 05a.

One Of ord'Mary ski:!! in the art would;easily and readily appreciate the myriad therapeutic benefit-; of sack an anb-O.Sa antibody, for example, as noted above. dm eoneentfaiton of eneelatwg 05 in human serum is very high, 'Rins. when introduced kuo a mammal, an anti-C5a antibody that is capable rd'siirmitaneousiy binding to two tmcleaved C5 molecules -would be rapidly inacrivated in the· molar excess of C5 and would -ken no longer be capable ol binding to- free €5a in the event of complement eehotiiOn;, -«s^^ -öfl high eoneeetrations and/or frequent u,l-taaist ratten of this type of anii-f'5« antibody would be necessary to, efieotb, ety intdbit C5a, In the 00-,0, that I; Is produces:, lit rommst. the awi-Cba antibody tiescribed herein that retains the ability to bind to free C5a, evert In a mola? excess xifbseleawed ffs; can tbps be ad-tbisisiereil to a human at a much lower dose and/or less hciicently than, mg,, an atitf-ClS: aoobody anh effeetwoly provide the same or greater inhibition ol'05¾ in tin.· human.

Accordingly, in yet another aspect, the disclosure features an isolated antibody e,>rtguLs'mg two antigen-binding sites, wherein each ainigcn-buKling shy independently can bind to tree human f. 5a ;hi.'e-a· or nuchom-d futman 05 thC’5}. wherein, in an aqueous solution Comprising: -;i) a plurality ol said antibodies and (ti) a mohr excess of hC5 as compared to the me do amount of the antigen-binding sites, at .,öc.^iailÏisrï:ui^i:«jtt-ditijts.il^ijpjbv^-ttostg.s M least 95,5,9¾ <;h,5, o":.t;7.5, or 9157/ % of mid pKnoHiy of ant;books bmd no more than one hf .5-msdeeuk:. i.c., no more ten 5% yf the antibodies are bieding two molecules of hCS at equilibrium;· hi another Kspoeb the disdostmo features an isolated antibody comprising two autigembifiduig sites, v>. herein each anngori-fcinding site independently can bind u> tree human C5u ihC fas or yneiea'. eg human C5 hC5 κ wherein, at equilibrium. ami ander ettysMogicM eoadSfimtio .¾ emapfisistg; f i) a #srf ily of said ;ndlb-idles and ;il i a molar excess ol bC5 na compared to the molar amount oi the antigen-biruhng site,' for antibodies e ;·.·,· icm-f Rs-'a oi ouu pie missy oi ennbodtes retain at leas· one antigen-binding cite mailable So bind dee Mbps. in another aspect, the disclosure feature;; an isolated antibody comprising two antigett-bindifig sites, wherein each endge-ohinding si;e mdenondentK can bind to tree human C5a (bib oat ot nnelcm ed human C 5 SHOP), wherein, at equilibrium ami :qnd#ii#jy§|df©ti0ti ddft’tótek in an aqueous solution comprising: ft) a plurality of said an dioxine· and fti i a molar excess of iiC5 as compared tie the molar amount id the •aótjg^|iJÖélb^tt-^'--(öbMtaboUies'h each antigers-binding site of ηo more than.5% of said iddraiity of asiïbqdsèy is:bound to a hG5 moleettlei in sotM.S'mbtM-utsgnfenïf-any'of the isolated smtibixises described herein, the molar e^eexy M:&amp;fi#ai a {&amp;g.ent least a 2~foki, 3-tbkl, 4-fpld, Mold, h-fold, "Mold, H-iokl, sMNId. oreren 10-fold) molar exeesx. in some embodiments of any of the isolated antibodies described herein, the physiological cm ids bon is 3 s> jsM NafkPO.ï, Mi mM Nra>HPO*. and 150 mM Nad, at pH · .0. in some embodiments ofiaoy of ihe isolated sMbqdies: described herein, ,^ii'-Mti.ph-ikéfhg.#«rt»^èpdh!3shl1y· can bind to free 'JsCSa witfra :¾ that is less tea i ,25x #·9 M, Isr some èmfeodimestrs, each: asti i gets-bintiing site cast independently bind tn free hC5;twith s suhsssnosnoSstr stbnssy s«ee ahovef

In some embodiments of any of the isolated antibodies described herein, >he Isolated imbbody composes c bob; chain polypeptide c^mps'lsing tbc amino sold segue no·.· depicted in Sf,Q ID NO'43 >e«,| a ban > chain polypeptide m-mprisiny the ammo ncid sequence depleted in Sh'tb TD NO:45.

In some embodiments of >my of the Isolated umib>xiies Jcscnhed herein the isolated antibody comprises' tit a light chain CTM<I con poising she amnio acid sequence depleted in She} ID NO:20' ni) a, light chain ('DSM Compos'lnn the amino steed sequence depicted In SRQ ID NO;2 1; tih} a light chan· f OR? comp! king the amino acid sequence depicted in SF.Q ;D NO;.w: ;"iv· a heavy i..bah; OOR! comprising ihc arm no acid seqstessee depicted bi SLQ iD NO:2H; {v} n hesn-y chain • f!jD|£|·tóiip#14-fffu^dibe:tlepieted·ia SEQ f,D 440:40: and $v$> a hca\y chain CDR2 cosnpriffiia the amino acid sequosKC depicted in SBQ sD'NÜ;4?. in ".one embodiments, she -scheed armhody can c-impose am <d ύ-c hght ehasn d s>R sen dcscnbed farcin. asw '4' 0w iiuhu'Uasn wsnahic <cymes described farcin to c . au\ ei'die hsn'nanwvd light chrnn \anabm regions* ,m\ ns she hems chatn OM< r>.w 0,0. pfa J fares·*, any of the Vum efatn variable regions described haren·; *,·.; y.. asp. os she humanmed he-o\ chans ur tabic mgsons h ot assy «nimble eombn-ations thescoi bee ess, Khdes i os 2, hs anothei aspect, she dis-Jos^e leesuren a medvd Fh treating a hsssn a* af'i'sscscij a ssh a c<mndcnscsr· associate j --njc; ft; g. a ΓΌ asyoes.tfed complement dfa-rdcr) sshesein she ns-jhed includes admisnstcrine So She human any of the fa-koed asffibcsbies ifa»rMii*ii· sufficient to treat the coinplefflestt^sspctMeil disorder in smotfa·· aspect, she diaciositre faaiures a method For treating a hitman aMieffid yd til a C S a^poetatgd èosnplósMnidtsbsidesy whet-eist die sfafhotf cótmpïtsffi admististeristg so she human--at least IhMe.g,, at least,0.7,.0,K, (Vi, or 4 x tng.of any oF she Ipeased antibodies described herein pet he b<ply weighs of the human so thereby partially os' completely bind arid semamie; ivanogmm levels of bee <.'5a ter greater than equal so, -or as leest Id ;e g., 13. 1-b in, lb, P, IS, id, .:0. 21. >2, u.v 24, or25; ant n. in another aspect, the dkcio.O3.re features a method For treating a human ainiemd with a Chawomemwd eontnlement disorder, wherein sin: method coissprisos adnurslstering so sfa human at hots; 10 mg s'imp: cl she isolated mssibodies described heretn {ter l:g body svemhr of the hitman to thereby-partially -orcompletely bind and \epoe,Oer mmogram levels o I' 11 ce 05a For as least 24 days.

Is; fetuaher aspens, the disclosure Feature a method for trentinii u hutrsan ssftllcss.d v, lib sj t'5:i-,s>Nuekted cc-nplctnes;; iioorder., wherein she method compose:-· adtnhhelcrnsu so the human anv os'tite ieelated aubbodsci- described hereto tor s'··;· cKitmpiv.- a pharruitceuiical eotvspo^lsls.a·! comprising any oi site i-n.dewJ ansthodica described hercbfF its ast aspeasni sul&amp;Iestl to fa) achisïvd molar Corn valuer equal to s;r less that· the phys-osug-e molar eoneerssratw··; oi uneieuved h05 wni th- jxtrfsuily or conqucteiy hw-d and eequesser sKsthoishysioiogtc Icsele of Free Γ sot. in some esïtbodiotenis of any oi the method.'' :d>.Ci'ibod herein. <s.st antibody is administered ίθ a subtecs ie.g., a human} in as? amount stnik sen! So achieve a nse-isis Omsk s?sloe that is subsusutUsily lover tM.it :tlt;gρ ityMOlsagte sPvd:&amp;r gMeemratisM Pf upbibaved -05 (g.g,l?f IJ},

In Md?d eiöbbditpbUtf of any of she methods dbsebbéd leresh,: ibe Oi»sk whte

&amp;,. ο.£.,:??ο greater titan §0 siM fór #f?ptrp&amp;Mai$%v &amp;$· I8^h||)s.· 1¾ some embodimssds,. Csnax levels are no greater sii;u; "0 -e.g . Kb >0. 40. ’0, or 20) nM. In some s-mhoJirnenN. the Cma.x value is no g-eafer than aggros ;m;>mk 100 nM is? sonte eiitbodink'nis. she Cnutx vah.se is s?o greaser slum 200 nM. In some embodiments oi anv otdhe methods described herche she Ctttas. value is,, c.a,, no greater titan 400 nM ior approxusttsseiy 4 mg kg ?. in some onibodbumus. sits.' Csn;t.\ value is no greater sintn: 400 se.g. 550, 300, 250, 2*00, 150. 100 90. SO, 70, 60. 50. 40. 50, 2«} or 10» nM ht srsodiet aspect. Ote disci ·*»?ν kamres a method ior treating a human at'isksed with a i'5:o.?ss--clued ounplcntetst disorder v. herent site metis a 1 comprises administering to the hsn?um ns?·. os'th·,. isolated ;*i?bhod:cs described boreu? sor tbr sample a ph*rmui.eutio;if cesup-;ibis<-u C'-mpsising any oi the isolated ^rush-dies #sedbed Iteredi}; nt un amount sultkkm so tat achieve nu den <. snn\ s «due·? equal to less than, substantially lo'ver slurs? the molar ph\sis>logle eonevninuion of tmdeaved l?C5 see if;· pari tally or completely btud and sequester rittnog'-arn levels id tree ('5a for el lea't 12 day,- ie.g., at least 24 days'? Suitable *. 'mas valueo ate drocsibod above.

In some od-ohi-vess .e'-un; ot she me» hods descnN'J besem site t's;· :?sv\ :.ee<l compknioui >hsci Jet can be, e ,., one selected Oont tise gsoup eousi.umg ot’.'Cp; I', ueuio respir oory do-uess .-ysub - 'me ; A0s Kt -epee shock am-pi? os plu'kpid usskhmme, 4 .uso.bopasi smst-pito-phi'lipid syndrome. ds-co"msrnen snfmv.i-vni.n e osgsiha;·m, hems m.-phsaps ·, -,.,,.'Opasime'-. S'.ndrems. bun? >>; son -.re bum m-brnKe IR I l P s\ miromc * ómmohonc .menu,? è hm used / o et eu. - ία-ο- and l on r’tjtekt eeunt'·, iutlatumanon-fUtiuiOsi pan?. * ‘he media kd uvussopmu.i, ago al.ued m mines begess;·. rasmn i -\\UV ehrmue ebsiruebw pmmon,,r- d.a-.ase, .md ?:l:S0S?ms?:S o id isrtl?rs Os. in \ ,-t .uionhes aspect, the dis-, h<,-um ieassne·· a eouuO'niot? eosnp-ioi'g α p!m nifv e.f isi.ilat-.d insist'tides esu'h usumo-b so'sije ptut-dsa es^epri-iing t\\., antigen' binding sites e.heimnt ea-..h .mimen PUninte. mm ntd-.pee.detniy can l-ntd re sV.e hnstus? s 'hi hC^atO' -aneiea\ed itntn.nt s °· Or 51 .os-l v\ lieten? sn site ρ*ο'Όηον i-fhumen C5 (hO?! -uni un-kr physudoeka: rmniuiosrs. no more shun 5--., of the asuibodk-s of she |·ίui-.ΐΐ!ï\ af <.qubb.)mun uompnoe fa,··; ysuigsm-hsadh-ig sites shmdnsnciassh bound to tmeieirved: IjC'Sv

In -s.fssre curio Honeon- oinsn -o'din oornpnssi-ons down'ined herein. she peso ejnsge ortho plurality in any pc rsie e Hr b i -of i n g u on rims m boo coo bo O'- sunned using high per ic-rusenee lipoid ehsosssci-'grapln HiPl. (k. In v»>me esubedisnesus, the phyM-iio^kai e> ouinnuo. in obi-els do.' son sin-dies me evalnased emnprfses she Ο'Πο'Λ lug e:oni»n><;-c inenboth-ït cl I)0 0 <e g. a ;ookr 000..0:-- te.g.. a ? -0dd nvlar -;'<Cna;-! of hC5; v-essh she plurality of antibodies at 4 °C for S4 hours in ;m aqueous sobdson coOii-rsoing 3.9 m.M frialkfÜ^ fief sttM arti 150 rtlBaGf at pi 17.0. For the purposes of this disclosure. the yoMtion issht&amp;fsied at 04 hours at #£: ds: ov-n.-ndcrcd io bo at equilibrium

Inkobie ombodimottfs ofiany i#itesosriposifions described herein, iso more titan 5¾ MtM antibodies of thé gfhraliiy comprise two antigen-binding sires SimtiteOeribsiy hóntsd nr nnolesved hC'5 under physiological conditions and in the presence m' as loan; a 2 tom «ml·» evens of hC? to sonib-adv.

In Ff the :a>mptmhk>asrd^cri^#d'-^fi%-ito mere shns; kb, -idhe antibodies of the plurality comprise two anUgen-btruling sites s smstl tmeoMly: boe ad tg: ttne fetrikt! .HQk mol ectdes :as eM foakfi asat g B fildk tolkm-ing mashsnku; >.·(' the pk-saldy os sussibodics wult h(. 5 as 4 X' fo; k-l h-assn. in an Upsteoas sbl utios comprising: 3J syiM WaEiPCIk,·: if I ®M Knri3 ®η and ISO isM NnCI, at pHkO. in smother aspect, she dkclossue feature:- a eosopoksiou eornosh-drso a plurality of i-anaied antibodies. each ssalb-ky ο l the pinna ip. comr-rnisn.: loo amiuaiobindmg snes., whereas cads ;uu-oosobmdis;u s-sn independently cstit bsisd so tree human C5a (IfCSktor unflekvs-d %tMi£ OS: {hC'S),. andl wfiereht hiy snore thasr $% of the asdiboslfeh. 0; she plurality comprise rn -..,- as digest binding sites rimuUaneoorly k.-usu'to nsu leaved IsC5 as emdtMed (&amp;.§., as fag HP L£.) fitlkiwMg IncuBatloa öf the jllutrifirinéf aötlbödièa Patfs :hC5 at: 4 *’€ ibr S4 höttsy in an agadpM splaiiots. ao:iftpf:kihg:;3:>0 tsM Nal i.iPO:, 0,1 mM Na;HP04. and 150 mM NaO, as pH‘7.0.

Is; yes ursotber ssspecs.. the diseiosutc fcamrcs t; cornjH>sitios; cnmprisisye; a 3>hsra!% of isolated gntlbisrltes, each :aniihiedy of the ptssraisty oontprisatga first SiStigost-bii-di-ig sss-e surd a second rursigen-bisuiing rise, vvlserds; each antigesvbinding sise independently can bind to- free human (kb; sbCaasyr smcl-aaved human Cb (hC5). wiierein c&amp;ch amigen-hindingsitc independestdy can bind ιο the tree hC5a ρ· dypeptide \van a K=> that is kss shan 1.25 x I O'* M. and wherein. in the presence :&amp;f

Immsp €5 (hCd); and ay byalaated high parforsnaneb liquid site of at least 95¾:öf fc ;:piui®lrty of antibodies art oeenpigdify yneleaved hCa-in the fel$o^4hg;<K|!htlgytit|0pv:<iSi iMti0-aftileis^ihlihg· site binds uncleaved ht'5 gad fie:^0h-44htig^htiklitt| site Ιό anbodralf or 0||.the:i|fM^^eii^imitnf.:'S:5ti· k tidbowad and tbaseeond aabgf ^Mhdirtg she Mi# aucleaved hC$,. ht yet another aspect. the disclosure fe&amp;mres a composition eompriamg a plurality of Palatal antibodies. each antibody >>f tPe plurality comprising two asvhgetj-binding sites. wherein each arntgen -blading she independently can bind to tree human C5a ihCiat oruneleaved human 1¾ <hC5). and therein, in the presence of human C5 the5) end a," evaluated ie.g.. using high performance liquid clu'omaiogtaphy {RPi.C'n andOirphysldlpgioai. böiidMeay,,. at least f5¾ of the antibodies ©€ the pfar&amp;fsty eontprisd at: feast ode tóti^èh^bjö^ihg^lth bapahlh Of bidding; to ffee; fcC5&amp;·;· ift some ebibodlmebts bf any of the compos Hid# described herein, die plurality of are do.-dies 0 evaluated in the presence of at least a 2-fold molar execs;- of hCT: antibody. In some embodiments of any of the compositions described herein, the plurality of sruilvidics is evaluated in the presence os'at least a 2-ibid molar excess oi hC5.antigen-bi ndu;g sites.

In yet another aspect, the disclosure features an ts eluted antibody comprising wheptn: the antibody binds to free €Sa or useleaved 05, add whepin one of^ip untlgeo-binding sites of iha aplibttdt'tenudns available to bind free C5a In the presence of a molar excess Pup at lenst or greater than a 2-fold. 5-foid, 10-ibid, 15 -Md,. or eeen a 20-#bJ molar excess) dl «oeieaved OS, in some embodiments, the antigen--binding sites have the same spociikhy te.g., the CDRs of each of the two antigen-binding sites share identical amino add sequences). it? some embodiment?., free. C5a is human €5a, In some embodiments, the antibody is croua-reachve herween human C5a and ('5a front a non-hunts» mammahan species. The ant-body can., in some embodiments, hind n> free Cva with a Mtbnaöomölas ailfoi ty; Id some euibodfmeu by ihe anfibedy has so affinity for G5a, that is at least ϊ'Οβ-· fold greater than its eorpspoitdiag afhiiilty for uhClcaved CS,

In another aspect, fte diselcsap fëatdps an two antigen -binding s;tes: wherein each antigen-binding site independently binds to free human Cfa i hCkq or unc heaved human C ' ih05 >. and t\ herein. at ;mv corseen-raiion >.-f ancleat cd ΙιΓΓ' : ear. hi a molar e<ces\ ofuneleaved 1-(35 du'i hC5u}. at least one of tie amtigen-fevnilmis .sMbS dfiig ghltst^ '$&amp;$·!&amp; Ü# %§§$,. (§>&amp;*. under bunco pnyssnogleal et-udinons. e g. in J inmost blood arsenm-i, in another a\pa-. die discf-mv leauuvr a composition eomorg log.; pluruihy oi isoluied a; urn-cues e isatcn each .mlibi-d·. o; the plurality composes two amsgen-binding si-ee. therein each ot'dv am-gen-btodsng sues m-kpendemlv mm hmd 10 five human s. ?u i hi mo or unekar .d human \ s i mm- ami therein at a molm ratio οί M s antibody 4(35 p no more lh«n, -r lav shun 5 re g , no mote sham, or km than 4 k 4 p, 4.", 4 6 4 5, i 4, 4,- 4 2, 4 k 4.0 ,V-> be, 2 λ 5 6. k5, .14, .< 4, 4 ;<. 2 5, 1.(1, 2 V, 0 s,.: ' Ιό 3 >.; 4. :?m : .:. 2.1, c.O sm. is,} ", kb. ι \ i.-i i k 1 .:. 1.5,01 j ·.»., 01 the ant η todies m'slse phuainy comprise pao antigen-lending m;ec> Mmulnnsconsh bound so m\ue-ivcd In 3. 1?- souk embodunems, each anisgcn-buKiine site Indepeiidemk can bind 10 tree hóón wbh a Ko that k k-sn Pom 1.23 x JO" M ior. for e>;ample, with submmomoiar aifirmy}. in another aspect, the disclosure features a composition compr-smg a plurality· of isolated amibodios. therein each antibody of (be pin rainy comprises two amigen-binding sires, wherein each of die tmtigco-hindmg shea independentis can bind to fret: human <35a -hC5ai or uviek-aeod human <35 <hC5s, and wherein. in the presence oi physiologic krek oi unekaved h<35. the antibodies of the plurality partially or eomplendy bind and w-quesCf iumogram levels of free ('5a iot greaser dam. equal to. or at Jeam 12 (eg., 15, 14, 15. .16. I", lb. lb, 20, 2i. 23, 32. 24. os-35'5 days when admimMcred to a human af ik-se- of I trig·kg or higher, in anodic; «speen ike div„ lo.sute ieumfen a ..ompo'Utlf'U comprising « pinrallis ut u,-.dated annbodse'·. 0 haem cued antibody id die pUimhw tviepritv- ioo m-ogen-binding sites v-hemm each -u du: antigen binding sue.··, independently atm bind n> tree human C5u - he'5at or mv :00(. ed human <35 i iiC'5 s, and \<- nereftt, ttt the presence ot pitysiologse let eo. o[ imeleawP 3<3'3. tbc auhbodies oi die plurality pa run i it er completely bind and sequester nanognim levels os fret ('5.3 h-r g tea it r dmn. eqnni so, tu leve id ic g,, 15, i4. i a, 1 e, J", IP, ; d, 2*4, 21,23, 2.\ 24, or 3''-1 days when administered so a issmnm -U sh-sOs of s*d mg. kg >-r htgiiet. in ajtothet «space, -he disclosure ieauuvs a eosupoou-m coieprcinp -i plurality of tsoi-tted a an bodies, e lumeet eaeis am:b<'-d\ 01 the plundut -..ornpri'Vs two amigen bmdnig dies, wheiesu e-.iCh of Use antigen-binding -lie? independently enn bind n> iiec tenaö: C5a fhCfSsi) ör tmoledy bd lipman CtS- iMl$), apd wtóiihy |ö the présesee of physdodogie levels of uocSouved hC5, the aniihodies ui the plurality partudlv or completely binds and sequesters nanogram level* of free C5a tb;· greaicf than, equal to, or at least i 1 te g i f <4, 15. U\ i \ Ib ih M,.? I. ? '· 1-1 ot 15) dnv s when ausrunistoted o duw-: aetucv tna mol o' t mas \ahk-s subsunnaih lower than the mokt? physiologic om;, οηΐίαίιοη of nnsleaied ha n

In smother tssport, the disclosure fi:asui\o n compoMPon comprising a plurality ot boiaioü -oo(i-.ijK'y ohci.m each am mob;·· o.·'tlv plurality comprises n\o innige»· buiding M?es. wherein each o! the <mnguv οοκίιηο sues indepemiontk tan bind na free human CSh ihCouf '>r undem ed human Oh < hi. 5) ,mb therein, in the presence of physiologic ko o Is of un<_ leaved hi'O the antibodies of the pkmdity pmtinlh or enmpletds lood end sequester piSilOphyμ<fume leiclo of free o'5a when admi-lists red b <l"Sc? achieving molar Otnu< values substantially km or ban be molar physiologic coneeniraoon of mm:kmco hCS.

In another an pec·, the disclosure features an isolated antibody composing a first antigen binding ok· Mb a second nmigmi binding ,bk\ wherein each antigen binding sib bdcpendebly can bind !< iwc bus bn: Cx: tSiOha; o? nnelensed human tfh : id'fh and w bemin o fen both amigeu-bitiding site' are t'nllv-e\\bp;cd untd, cm... onder human physiologic*! >.ondbions. mg, m kimum blood or serum!, be following bimling configurations ure possible «i ' the fust antigen-binding site binds bee hf'ha nnd the "-bond anfigeu-bnidlug site binds unc leaved hi x di) she lust anugendaindtng site bonds free hi'ba anti the second antigen-bonnon. site binds free 1-05:).: ortitii the fit si mingon-bi tiding site binds tun. leaved hi."? and die second antigen-binding she binds free hOis. in yet amatlier aspeek the dtsciföpt^.-fe^tötëts··#» isolated mtilbqdy cemprlsiu g. a first an;:gcmb:mn:w site and a second antigen- binding sit:.·, t herein -.01011 antigenbinding site independently can bind to free human ('5a thf'óai m otileaved human CS ilk. 5). wherein each antigen-binding site htdepeitikiith can bind to free hC5a with a K; > I ha: is k-ss- than 1..15 \ Iff ‘ M tor. for oxunple, ul-h xOmmiomcfu; affinity t. and wherein, in a pltysiidogk.it! solution eoiPaining a plurality 0: the antibodies, at I vast §50» of flip: dtikbodl es aye lb flip following eonflguratiofisi (if if te fii'st arii%gB«M ndf ng Mtc bind'.- free h('5a and the second antigen-binding sue binds uncleaved hCr \iij the first ant:gend>itiding site binds free hi'on and the second antigen--binding sue binds tree tit'5a: tint the fust antigen-binding sue binds urn,leaved Inf $ and the second imtigen-binding ρΐΐο bui;ia tree hCsn; >; s i the ;rs» .umgcw-bindiny site bittds imekavcd hC5 cmd ï-hs. second antigen-binding site is unbound; rvV tbs first amigen-=b'd i-ïi-g; JiiC-Ssb^iisi^ $tg-hèdiö»4j f»$&amp;m*ht»d'mg site k unbound; {vit the first smigciddndhig site i-; unbound and du: second mui gen -binding. «de binds undmused &amp;£Si(yH) ihp-Srsf $iitigen^ihdih§iand the second smtigcn-bmdmg site binds inbiu; and {vin} the first ;euseen-binding site is unbound and the second nmlgun-bhiding site is unbound in another ;.;sg:.-ee -be disclosum feumres an h-oSated antibody comprising to v imdgea-binding sites, wiierwn each antigen binding site inbcpo'klcmh can bind ;o tree human C5a ihf xs; or-.lucictvai human h'5 dtC5i, and wherein, in a mobr esecss eb' .nn.leaved in? 5 o'-er hC5u, die antibody inhibhs by m hu-c dirdi hCmodurxmdent hbmM rfeteitiphiT actiysddn at;a tnokr raiir> ot':i, t omheen binding sin;. hCba's in some cmbodimenis of any ot me am i boe les desemen hemm, die .i^Mg|iï^tïbii%'tó'|ii<^^rbfei«tidèrtbrimaïi:-phys:i0h'gK'ul conditions with fulK -toldeJ. native, human Cèa and O protein- in some embodiments -o'any of the antibodies dc-enh^a heren'», the antibody binds to free K.oa with a Κ.». that is less than 1.25 \ 10'' M sor, fen example w idi subna non to iar affi η i ty s.

In vet soother Mpeebdlïcfeciosurc features :m antibody that on bntils to free a subsm no mob tr laffmhv -and <bi binds to tree ( 5a wait an •«®hltyth$£i$df:Iès$i H)0 te.;.·., m lenst Πθ, \.70. let*. 140. 150. '-off ί "O. ISO. i»H\ 2¾ 21¾25¾.:1:75,3M, -too. eOO. «Μ. "tR atR <>m. iOOt), }m\ MK>. 4000. S000, 6(.HR "1)00. StfOO, 90iR or ERiO -fold greater that» its corresponding aibnuy for uiieipved: C5 protein. In a Cdrpptis&amp;tg apliirafit f of tie antibodies, for at icsst RR of the -antibodies. only one anhgcn-binding site of the antibody binds to uneleawd 05 protein, wher-ms the second aotigi.-;»-binding site remains available to bind to nee 05a {The iiOR cm h.ne die monk· acid «cqueece depicted: in SEQ 4b) M>: 1.4

In; Mother Mpeet tlm diRlosRO features: a metlod for iteatoig almftmo afflicted with a 053·associated complement disorder, the method comprising Odmittis tering to the frtrsffin tr eoteposidee eomprkirtg, a plgraUtv of Isolated afrtibodies, whereirr eseft ontibody of the pittmlity gornpnses two an dgen--lind:iH:g sites, wherein each of the antigen-binding sites independently can bind to free human ÖS» (h€5a) or «cleaved heemst :GS fh€5|, whereitr at least: f mg. of the #öiiftottres pér kg body weighs of the human ;s üdm’mKiorcd n> the hssinxu. and whereisi sdmlnMirsilon of the ahtlboiiék is effective to partlafly er edaïsptetely bind santt s'eqssexicr nanosram IcucN cd free (.'5a ie? et lesss- 12 days, iss another aspect, slu: dssclosn-e ioaiurea a method Γ*>r srea-htg a hsssnns: affheied 'o ,'sh a co mn; e so en x ass >e i ;de : i disorder { e.p.,, a k οη-.χχχ tek-sted complement disorder), du: method onoprisisx: adofsaxierme te the hssmusi a e<nTspossiuu: eim-prising s plurality cl’iv dated antibodies. \i herein each antibody of the pUsrafru ircrapii-'v-'s Ολο arsticess-bhxlistg s;:os. wherein ends of the aoisqesvi'mxfnsy sstes osdependesuk· eau bmd to bee hitman ('5-i thC5*i: or utscleaved human O thC5). whereisi at least p) π sa of the antibodies per kg both weighs of site human is administered to. she human, as.ai wherein adsusnistrmiuss of the sadlbodies is effective s<' pasted tv or comp lately bind artd sequoutet rsanogrum levels of tree C'5a fur at lease do das\\ hi smother w·· pec:, she disclosure teannvx ?: method for treating ?i human siftsicted m Uh ;s compleotenl-saa^ocsatcd disorder te.g., a C5&amp;-associated cotηρΙ$$ί$ά£ disorders, the method eomprisstsg adtninisscrusg so the human &amp; cornposiftou ebbipfssrtsg a phrrslitji of sspIgSpil ant sbödias, whet'dift tooit issilfedy of the plura thy comprises two erdtg',nxhissdmg sites, whom in each of she ;mfsgess'bindh'ig sites mdependemiy can bind so free human C5;s (hC5a) or umleaved human C5 (h(.'5s, wheresss she annbodles axe administered as a dose auffieiesn 10. sat achieve molar Cm ax \ sslues suhsinntiaHy imoves' than the rssohsr ph> sioiogie concentration of UHiteaied hf.'5 and tb) partially or Completely bind and sequester nanu^ram k-yblè-^f free 05 a ibsat icasi 12 days. bs anotiuN aspect, she dsvJosose ietiunes a rnedvd lbs treaong ,s Isusn-a? atYiseted wjth ,s complement-as-ommcd disorder uw,, ,s s.' 5a-!-,secxiteb oontplemem disorder', she mu hod comprising udsinnwersis,.· to she hu sixes a ι omm»' sihm e>apprising a plus::.::1) o·’isolated .intiK'-Jsc-x v,heoesn ex. is .: η k bod > sa'sis·; pksralsis cs'Se-pnae,- two antigen hsxhtsg w here::; e seis of the anitgeu-tnndsug "te-mdepindenth :.an hmd so fses. human s fxs sin. 'xs' os wx'kased hsssixsit k 5 shi'5> ο hereist the amsbedsca are adstiinistcwd ;d a Jose sufficient d>; sa s stehics»; Osola:'

Csivalue:' uubssaniksiK lower thint she mokn' plsyssofogsc coneeufration nf imeteavcsf fsC5 sM (b; pattsaliy or complekefy biniJ and sequester aasiogmm levels of free (’5a s’·h' as least 2-4 days

Sn another m-pcee, the di'Closure tcunnes .o. method lor treating ,-i human o!lllcn.-d obh ;s s.om|domuH-:i<-soeutted disorder teg., n C5a -assoeiakd coropiement disorders the t nabot! comprising administering to the human a composition comprising a phumbv of isolated notihodies c Pof cm such ;mdh<Cv of die plurality· eotnpnsCN h\ v> umiggu-bimlmg -des, o herent ea«.h td'the hrtyc«·b.indm§.sites· independent i\ san huid io free human Cèa fhC5,f oi nuelcaved human C5 (hi'5;, 'therein if:, nmihod-es a;e a; lm in': si»..-red. m ;· dose sufficient to: tat achmse m< far Orotts s aloes subse-mtuf-ly f av.r time ïhe rn-dur physiologic concentration of uiideCHed hC5 end (irn partially -r Completely bind and ,>cotms kt p;-d hop'd y s i cl o g:c levels >/f uee k'5a

It is understood bun any ui The mnupokhom; tem,, sompr.lsing a plurality of a?H-bodies >'!}' isolated ;-upib-;dies any., that rendu, in the presence of 05 or run lm exeess of C è. ;> [ree I nb :nan capable oi' Pi mime u,· Uee ('5a; described herein can he. (M for mutated as .pharmaeuulsca!. composto tm v m acebydabey W&amp;b: the d! sciMiiM. {hi) Included -η ?hcmpcm>o firs mesenbed herein s or tei uidaded in the ere-idled syringes described herein. Λχ describe** in die '.unking e\atr:p!es provided Iteteui, the inseutors have also beteyf fdatpot only hlpdc with high aximity (.subnain.-inoinr aft mi Is ) m fVee hCW, but ai concentra-i-mr in excess oi imeleaved C5 also in in hi o. terminal eompk merit complex ;T(. Cl ibnmdlon in a dose dependent manner, even at coucemrauoos oi die emi-Con smdbodv h- greater than h.5-j<>!d excess of C5, however, mhihimm oi TCC is no? eompleie. While the disclosure is by Ho means limbed by -any pameuUtr theory or mechanism oi action, the antibody may inhibit TCC iornuitiun by binding to ;-n least a (me; ion of undented C5 and preyemTng its cleavage andfof otherwis e pmvsibihg: the success!id sssocMic# of C5 tviih additional TCC components. The m sent ore appreciated that such an am-body is useful hat rummy complenmot-moociateb disorders, c.g., m ohied f m·: plays a significant role and the CTmeomCmoy TCC may play a lose substantial role Such disorders can me hole, e.g.. seps-s. auric respiratory distress syndrome tARDSt. septic shock a mi-phospholipid syndrome, catastrophic anti-phospholipid urn'romc, disseminated ini m t a sc u ki r co a gt s k: t i os i. lupus nephritis, Goodpasture's Syndrome, burn si? severe burn, asthma. Held. P syndrome f/-?enivSyPe anonum i.Teuued .fiver en/yrnc? and /mv f'P-aeicr eeinu;.. intl-immadon--t-duced pain. On-mediated neniropen-a. age-rekted msouiar degeneration t AMD), chronic obstructive puboosw!'} disease, and rheumainid xrihritfö, the inventors aM> appreeuted hut use of'such es ,tmï-05 a ins?iboJ\ te nam thess conditions. .ηηοηρ ethers, mus pi - >\the an es,.?; mere beneficia; wtk'fx prefik- :e compmed η> esc o* totsmnui eompiomont utlnbuer, drugs Λλ noted uhoxe ene jtouhk ^omeoueeee ,>t mbtimhig terminal e'-inplctneus eompw mms such tu l 5, t 'O 0\ O ". * V e; O' :;· dccmse-cd promonen bx the hev itewnne ^ys-esa agwmt the .nchpealöi-'d baefona that ; -rrasnal e· -mpl;·men* ordinahk Ksos Sar o sample xVs thé antf-OSa aikifeddies desctibed u; hu,' :-,.eb,in ah-da; she (' 'u-tneduted millennia top, iesjvn.se, ί»ιη d<> nes eomptevh bhehb th„' tonmeten al she tortnma! complement cwnpkx hut bsc-i these ettvtpsulmvd OiCicru, >ueen?,'> tocetMng a fhentpetme anti f ;u antibody desenbed Ik rem mav ma ycoem. nateeM v uccnutiom e y . <t xaeonution against V v,s<r(4f t%ëtii-!igU0è$· add Mksahdtz 1?hrtiMt&amp;i»Bs'ü0n of the TCXh white sof wholly akrogumtg is-rminal complement's unft-rmerubttf! response, may tst kto; reduce TC C induced htikur<mniiOn as i issue injury, Ί he partial '1 (.X' inhibition, in combso-ation o ah inhibition ofCea, is ixTtexed ίο make the amt-Chn antibody an even more potent anti -mih.imm;nofy eotnpotmd

Accordingly. as anmhor aspem. the disclosure features an antibody oramleen binding fragment thereof that binds to free Aba, wherein the free (Aa is human *.5a ha\ tny the amino acts! v-goera. e depicted irs S?i.O !P NO. i, wherein the antibods inhibits the binding ot 05« to 05« roeepvr, tmd \xherein the antibody parhnlk inhibits formation oidhe terminal complement complex hiVO, Pur da; inhibition by ms ausi-C Xs antibody or mnlyen-bindwg fragment iheveol described herein cast be, e.g.. a complement setts it) that is. ist the presence of the annbosiy, up w.vr!K- greater titan Ml ;e.g.. 75, ?0, b5, bO.. 55. 5b. 45, 40, 35. 50. or 25? 'b, of the cempfernetn aettvny in the sb&amp;estce of the asthbody. or a:tki gett-%ibdltt|i fragment tliereoi, in some: crnlv>dlments. the antibody or anitgeo-bindiog fragtïiettt thereof bistds so free C' 5a (g,g,, fret ftC5a) xcitlf. a: sribrisnontbiar affinity,. In:/some ótnbödimentsothê asübMf or aniigeti-binding iVagmctu thereof has :m a.fhnhy fo;· bee C5a that is at ieast ) Ob-fold greater than the corresponding affinity of the antibody or tirstigen-bitiding fragment farspneleayed G5, In aome tmljoifi fnen ts,: thy apt ibody or an bgeft-bfnding ftsgntent thereof inhibits by at iese? 50ι:·<ϊ Sorntation offCC as •..oncs’iitratiosts exceeding 200 fe.g., 210. 220, 250, 240, 250, 260; 270. ?40. ·?«(». 300, 320, v40. 5bt), 58*0, or 400 or naad μρ-tuL as measured itsuip u Of :50eq assay, 1« some embodiments, she tumbody or anacen-l>; no in:;, itaptnetr· thetcoi inhibits by ai least :¾% classical kumpiemcut pathway nedvutton a; eeruvafradons exceedktp 200 (n.g., 2)0, 220, 2.20, 2 t0, 250, NO, 2”0. 2h0, 200, 500, 220. 2-10, 2ó0, 3HO, Of 400 or more) jag/mL as iUOu.-ttSxd stship 2;- Vs ie-i«K’'' l'I.VIi,:! SV.-h'A <5 s ( ('tnpionietit 'Na ,t,s best tUkd SP tb,' w orkistp exttmp Ie»; in ;utotSk: .apv', όκ di vlosn-o je tintos a srvb'a O fo> ouksna a hunkin «ffVhx! wüij a ,O:'kp'om.'iH-,-s-'ii.Mk'0 disorder ten’,. a i Ne.;-sec iaioe iompieavau disotber of a eompOrrvaf-tss·· -i nee,: utflanor>.tfery de .rbo* > 1 2c ntetitod ut-, Osje: ,sük\fii'totiiii; t·· the huftt-tu m οίΥνοχν. amount of en enuK>d\ o? v.necn bmdtnp haatneuf theKkt’iiixït ndnbib. Ou. aasdinx; of 05a v 05o ονορΐο: o,ud oJvtent fik anièVib, partial :\ inhibit' to.Oixtton ot' tb-, minimal Cfitnpk'nvnt complex i H'Ct Nee abox e i h. Ίο'· ···' Oi i ,ut tv aa\ o? those Knots n ut th- jr; or 2·; vnhcü is rent

in αηοιίκτ aspect, ik disclosure ie-dutcs au isolated antibody or sou peen binding ikicaivia vevoS that hind-» tea hu ras a 0>a ivix pepode huvu" the aneno «o ;d .'Cijucaco OpuO ta X s .0 0,; Ni f {. ba; doos a< a hind to tbc alpha ,. lad a ot' uuciose, cd, aai o.·. 05, wik-reut the antibody ,η-neen binding Seigmcnf fhotvoi'lands to the lantka; ( 'N poh peptide v,uh a K;. that ts loss tjtae \ 21 \ 0; M in nnoiher aspect, Tho diseSosum features an is·dated antibody o- a-dig, n-binding teiptncut Νον-Υ that buide soa luim-ut t 5a polypepbde huvng the annno acki seiiitetkk da rad cd tu iV'O 1Π Ni.)' i. but does n<<t fund V the alpha s, ha ia of nnoieavad and' o Cs, ohevsa dk onuVnb ndiihit- by ut ievo 50d, husuan f' Nodependent human rvauvphii a-,te,mav at a molar rau-> of 1 i ;amipca bmdtng site 05a t <u some embodiments, ihe antibody tnlnbtïs by af se-td CO!\> butaan C5a dependent h-nnac nevbivphh aapt af as?: la aa assay In o ba Js 0,4 n'O o! aanbod; is used i'.s inhibit the !ic;>;sx.spibb:xostx;ntoe achsbx >4',' ;ΛΙ home o ('5a v >ksuiKJ in i's.sTripie s in so ra a ^n-hodsmeras. the aaidv-dy doos not οοηρνα,ο exetnpl.-try i'i>k panhie 5 depteted m luide i ba s.unc etnbodnuems, the autibodp uot in i^sasc onibi'dimonis, au n-oducJ a:ah.«-d\ ot aaapca -iiatdrap bapsnea? fherestf described hetein (htuk te u Staaata t.'óa pdspopod·. Stat tap tito .onmo aca.i seokicru.v dopata.d tn N{\" f|) Ν;,):2 lts si,snk ctuboditnetifs. au -sodacd ad;body or atuiyon baniv-p trapiaon: te.oiC'sf' dess rtbcil heroin eompttses a ftpjtt chata tvb, pcotido r ontpr-saty a Üeht oh-on O H<! comprising tik amino m.so 'sequentie depicted in SE 0 fD NO-30: a Eg hl tJmm tEDRE e<mtpreono the nmbm uutd sequence depicted m NEC5 ID N< >.3 i; :md n light chain QDR.3 composing the uïiïiïs» -.tend 'comence depicted in XFQ fD NO, 3 3, in "ene embodiments, n't isolated maim 'ij\ or anti gen-binding imgtnesd utereol described hoorn comprises ; tight citmn poNpepmic e>nT>prieuig· a heh? Jtdi; ('DRi compnsntg the mntit'i acid ssquence deptcied m Si..Q ID NQgNg a Echt chain t3f.'>k3 comprising the temn-f amd v.opkmoe depicted in Si'Q SD Nt Q’NS sod ή in. Et ehatn 0.)k 5 composing -he ammo a ooi ?;.-qyoitee EeptmeE in ήEO Hi Nv> ‘'3 in some einbvdHücnïs, «n isolated nnUhoih or antigen 1 End mg Ragt neut ibc'wi desct il<ed he mm mmtpfRos a he-sm chain po!\ peptide comprising. u heary eh;on < 'DRi o-imposing tne undue uetd sequence depicted in Si:O li> N0.3E n imm \ dime; CDR3 eompnmng she amitio a cel sequence depicted m Sis'3 iD NÜ;3N, und &amp; homm-, chain 0DK3 emrspnsmg dit amiuo acid zeeptonen depleted in SEQ )D NO.30. in some mobodkoemv. an isolmed muibody er unUgett-binding fragment fbC'k';»i dtï:f,r:b<.ü in.-rent comprises a hcuv\ chain poiypvpttde comprising' a hemer i/ham CDR! comprising Uk" amino acid sequence depte led in SdO ]D NO. .O: a heavy chain CDR3 comprising the umiuo acid sequence depicted in SEQ ID NOi>7; and a heavy chain CDR3 comprisiag the amine acid sequence depicted in SEQ iD NQ:30. in seine embodiments, an amEued asttibody :.!' antigen·binding fragntem thereof described herein comprises a hesny chant polypeptide c-otnpridng; a he;t\g chain CQR i comprising the ammo acid sequence depicted in SEQ ID NONE: a heavy chain ODE3 comprising the ammo acid m-qmmee depicted in SEQ if) NO:-Ira and a hear y chain i. DR.?· comprising the ammo acid sequence depicted in SEQ iD NO;4?. hioa.ime emhodinietus, ms isolated antibody of asuigeu-biskHug {moment thereot described heron comprises a light eiiam j>.dvpepnde comprieiop the amino acid sequence depicted in SEQ ID ΝΟ;·Λ7·ογ SE(> if.) NOOp. ht some embodiiftettis. an ROarcd amibt>dy e msPgombmbmq fmgmem titereof described herent eotr>prtses a. heavy chain polypeptide c mop nr. mg the amino acid sequence depicted hi SEQ ID NOS.:" or SEN ;D NO:33 M aotne èihbódittïcais, an Nolkied .antibody or aotigoikbisidlng' i'aOatettt: thereof tleseriVtcd herein comprises a light chain podypephdc comprising the amine acid sequence depicted tit SEQ 10 NO:3"; ami a hem s chain polypeptide comprising the amim mod sequemte depicted in SEQ ID Not33". 1» some -.-mfcodmvnts. an tsolafed antibody or antigen -bOOne fra genen; light èhain fxdypcptide comprising the arouv ;xhd sequence depDiPsi m PLO fD NO 3 b uud a hea\> chain poiypepnde comprising PK> nmim'· ac;d sequence depicted in SKI ID Nt !;33. in roe embodiment^ an h Dated smibodv er am: gen--Onding DigmeO thereof described herent comprises a light cintm polypeptide comprising the ammo·· acid sequence iienicf.cn in SEQ ID NO: O or SEO ID NO: I κ in some cn-boijin'M-:n.\. as\ isobieb antibody or antigen-binding fragment thereof decenhod herein emnpfises a heavy chain polypeptide Co mens mg -he ammo acid sequence depiCied ir» SBQ ID NOD? or SB\> ID NODb. in some embodiments, ms isolated antibody of asuigeu-blndiug bagmen? thereof described herein comprises a is gin chain polypeptide comprising fhe ammo acid sequence depicted in SEQ fD MON0 and a heavy chain polypeptide comprlsrsigv the amine acid sequence depicted in SCQ fD NO.2?. in some en-bodinvnis. an isolated antibody or antigen -bind-ng fragment thereof’described herein composes a light chain polypeptide comprising she aroirto acid se£|üened depipfei in S1Q ID N03l'3: a?ïi| a heavy ehain polypeptide comprising die am ism acid 'Sequence depicted in SEQ !D NO;25.

In seme embodiments, an vadated antibody or nntigen-bindmg fragment the roof described herent comprises a light chain polypeptide comprising die amino'· acid sequence depicted in SEQ ID NO;42 or SEO ID NO:40 in some -.mbodmvrsts. an isolated uniibody or antigen-binding fragment thereof dèscfifeed herein epipprisèyb: heavy chakt polypeptide Compriying the amlnp acid sequence depicted in Ship ID NO..:"·' or SB? ID NO:33.

In some isolated antibody or nntigen-bindtng, fragment: thereof described herein comprises a light chain polypepi-d-v romps Ming the amino acts! sequence depicted In MEQ fD NO;-i2 and a. heavy chain polypeptide comprising the amino acid sequence depicted in S't'.Q ID NOD? fn some eo-hodm-ents. an isolated antibody or antigen bind-ng tragenen; thefêrjf dercrlfaed hereiP: comprises a. light chsid pPlypeprtde epfKpflSing die: amino aeid sop >is'nee depicted m SKQ ID NO-phi and a heavy chain polypeptide pomp rising the amino acto 'X-questce depie km in ,0:0 ID NO’33. 1st some embodiment;, an sre-fated antibody -r aungen-b-tklmg {ragna to thereof described heven· comprises a hght eham polypeptide coniprisiug the' annuo acid sequence depicted in Sr O ΙΠ \0:Ï7 «nd « heavy cd a irs polypeptide comprising the nmmo steal saptc-nce depicted 'm SLQ iD NÜ:.v\ ht some embodiments un isolated antibody or amiceudoiuü;te. hum neut UiCU.au described herein aurupru-tu ·>; Item y chain polypeptide comprising the ammo si'kl satne.nee deputed In tiny one oi S c) iD NO:4.\ SF.O ID la 0:11. or Sc I) Ul> .NO;#,.

In some eiOxulheeuts. an isolated antibody or antigen -binding fragment ' 4ès#i(bé4·'héféiöi &amp; l|^tt:öfe.aj.jj.-jKi.!.ypeptide€ompïi5Óng the amino

Sp0' tï>Mö;ïb and a heavy chain polypeptide compiling rhe ammo acid sequence depicted in SF.O ID KO;45, in: some gr#!# imegtg, an isol ated an d h« >dv of ant i gc s-bin Jo nr 1 s *gt n e- h thereof described herein comprises a bOn chant polypeptide comprising ?he amino acid sequence depleted in St# fD MO; 17 and a heavy eiamt polypeptide comprising.; the amino acid sequence depicted nt SDQ ID NO;44.

In some ;.-mhodin\ nt\. an isolated aritbody or amlgcn-binding hugmetu the^eoi Jo·u ;><so U'.soln i.vm;u;- n #[> -John polypepoJe comprising she amitto .text setpiCtR'e depicted a; of Ο ΙΠ Net \" attd a hca\> Jut in polvpeptxic comprtsmg the smltm aetd ‘'equenee depleiea in SrO ID NON'g

In some embodiment'% a>-l.ucd mmbudy -.-r mtigsn binding ΐΥοοηχη; thetcof desorbed herestt coo guises a light chain polypeptide compttsmg site ammo no id sequence Jepk ted in SI# ID MOD ' or SFO li> NO. Mo

In some omho.jinxms. an -sphied jniibods or amleen -binding fragment ihcreoi beset ;bed betem smttrnxr· a hear v chub; pole peptide comprising the aufrut -(ten.· sequence deputed In Si.O ID N< b# e-r SCO' ID Nit:44. in -ome embodiments, an belated amdnuix or antigcmbtndhtg iV·· cut noth thereof described herein -comprises a tight chain polypeptide compr?«mg-the-am'mo aeid aequenee depicted in SFQ ID NO:.47 and a heavy chain polypeptide comprising the amino acid sequence depicted in SFQ ID NO:45.

In some embodiments, an isolated antibody or antigen-binding thtgmem dÉerêcjf described hsreih: eemprises &amp; Mglit chMti polypeptide eöinprislnir die stiiuiro acid sequence depicted in Shi) ID NO;36 arid a heavy chain polypeptide comprising the amino acid «qttence bepMed in SiFQ ID ;N0<4§, hi so Of.' onhodhiknis. an ;soia.k'd arndxkh or ;®ύροη ·ί·όχί:ηχ frays noot there® 'i-.-'U ;l«ti jietom eoxokO a Ilyin ι ham poh pepoie οοηριχη® ho* amino acid sequence depicted if; οί O fp NO 42 o*· SifO IP \O:40 hi -'imo eobedenoos, ni a-elated andboh or asnigen-biSidot© ioeioesb tfiercol described ho;on comprises n ne,ny chair; ppk peptide coupnen® the mxln© acid sequence depicted ® Slip IP \p, If m s£<\' ?D NO 4'·), hi some xiobolheenis. an odaied uifhfxxK or tmueeu-hnidnag fragment there® Os; pK.© iktvm e-imprOeo a hoin ; ham pohpeptide ceenpiODy ihc urn in-* acid sequence Opvoed m SeQ U) sO 42 -od a hem) chain peh peptide e<«nprism;i the amino ,k;4 soucskc depleted in O::0 ID NO,45, in ome eobedioeen·., m eolated undNah of ant-gen-bindο<: h-ome® ihooof doxadvd hou·® oomp!<se> ,-, light chaen poiypephtk eomprisiu&amp; rhe ammo nod -eouenec Ooi noted m 'Lp ID \0:40 and a heavy olunn polypeptide ύοηηϋ&amp;ίβ£: the ammo acid sequoxe cos>vkki in ShQ ID 00:-49.

hi Nome v-inl'-ti-hon.rns. an Related arOho.h or an;®®; -bind me. frays; out Φοχ,Ο Ο®"® U® hoen; m>d4 m hi ;0 e uh K:. hi® is loss than ~ ,\ !Ol"M hi feme on ;ho dime lit;- an Isolated mu’d«xh or ash; yea®; s; el icy ham neut thereol describes; heroin rends to hi On w iih η K;, that 0 ic® ihran O 10 1' hi.

In s-,>mc eiobodireeo;!'-, an lOmed aanbed} o' antigen binding fra on con thereof described heren; binds io Is* '5a u iih a iv- thu? is less Hum 5 \ 19 u' M,

In some emhodimems. an keburd mOho® or andycu -bimjmy ffaysnetn fhoi'ssd desciih-.d here® bind4' η hi 5-i v nil « K; , that is loss than 2 5 a PD 1' M.

hi some on Oesl is nee ;s an isolated ;mhb®q\ os ,m si yea-b; siding hng* neut ihereol dosorihod heroin hinds a - In, >.; p eh a ®r, that I- loss man I 5 < 10 M hi "omo esnbmlisnenss, ms Isolated andbrah or asn;yc;;®mdh;g d'-eusc® Uk-reof described hetem funds so I®'bo wish a Kj> -hut k, less ihran 1.0 \ U) *'* M In some ombodsme.nm. an isolated antibod} e anf;ge;i-b;ndme haymem shoreo!" described Sieren; binds to hi. da kid; a K.:- th<*i -s loss tie-in 4.0 s id 1' O

In some emhodiments. an antibody -nhihiis hy as ionss Ό o.y , .a Oak "5, hO. 55, '·Ό. or 95 of arcniOP ">' innnan 05;;·dependent human neuhopih) aciis ;ηί>·η Ό a rti'da." ruth· et l· s ;anhyend'diidirsy sito t'oa'e in s-,όκ oinbodlrneink, she muibody docs no eoniprOe e\osrspktr> t'DR pairing r depicted in I'aSpSo S in sone omSxjdn'ne.niS. the antibod} is no BN O'' I.

Sn \ m another aspect the dim Ktaure temttrer an oolmed -antibody or antigen bnidiug bi-mmenl fhetcof’ that cmnpnr.es a light chain ('DU set m set forth in Tabic a er Ta ice 7, t\>r example. Oe Notated and Nmy or atuigen-bitidutg. iraemem Ut eren Γ can cu-opcNo a light chain p>>lv peptide ootnprNhig: t i > a ligjü chain ODR; comprising ihearoin-aud -equenre depicted in ShQ iD NO: i Uk a HO·; chain ('Uk-' comprtsittg the amine aetd sequence depicted in SEO 1D NOOó; and a tight chain ODRn evtoprlOmr the a nano acid sequence depicted in Si: O ID NOtl +.?: (it ; a light a h;nn ODR1 cem;pr;smy tin.· amine a·:.id sequence depicted 0 ShO R) Nee !5b: a light chain COR7 coniprtslng. the η mini.' *ud sequence dCjvictcC in SEC,) ID NO. 1 ο"; ami a %ht uitam i..'i >Ro emnmisee. the ntnitn- -.and sequence depicted m Sb'O ID NO. i Ό, t lh; a in.-ln chain ODk 1 eoinprisutg the mum- < acid sconem...:· depicted m >d'.Q EN< a ItO. a light chant CDR.? cmnpnsmg the momc icid sequence depicted m SL'O Π-'-5 V k ib5: and a baht chain RDRo c< mipslsmg the «mine acid sequence depicted m Si 0 ID W ior· io ΐ i light chant f DR \ cmiiprlslng the amine acid sequence depicted ?n Sh 0 d) NO i " \ a lea in chant hd ik’’ c· -rnpnMny the a sin re i ;tc :d 'teqnence dcpscmd tu St\> ID Nt) S "Λ -usd ·! hgin chain «'DRè cmnps isum, the am me non] sequence depicted it» SEO ID N't » ; “4 t O a tight chain ( DR I cvinpi^nip >he annno acid -equeticc departed in O'O ID NO.sd. a light chain t DRd ..mopi'ente. the annn-t aetd sequence denoted nt Si O iD Ntv\\ .md a light chant h'DRa comprising tlte armsm amd mduoiice do-ncted m SI'O (D NOOm m I!hgh? cit.ee ODR1 compt irony the mnhui aud sequeitea depicted In NRO ;|a \t OO; a light chum ODRE comprising the amlrio ,u:ld sequence depicted in SFD ID NO. CD uud a hgiu chain OaSa comprising the mntne aetd sentience «k>n<dcd in STQ ID NO Do, t\iN a licht cinins ODRi conis't't'di!? dn. amhi-f ttetd --acutetc,.c depicted in SbO R) Nt) nd. a inde chain tsT>R7 cottipne'nt;.’. ilk mm tie .u td -eqneitce depicted hi STO II) N<‘> n’t; and a Rain chain h 1.);·:3 ecanp!nang the ammo acid vqitene*. depicted m SI R> ID NO'-hh, m. nt' a hgin ch»m c DR 1 c-amorc-mia the amino acid -.eqnaccc depicted to SL\t ID \0.m\ n light ehatri I DR7 uoipmuo the ammo aeal acipncncx depicted nt Si O ID Νί) *1ι\ and a light eitatst t. DU.1 ccmtpre ing the amino oewi \mnicncc detateted tn Si.pt H) ROD7' ttx) a light chain CDR t contpt Im-tg the ammo acid NOqttcnce detneted tn Sc D ID NO E'E a light chant t."DR.) cvmpnvmg the atnino acid aocinenoe depicted ttt SI >) II > \0 100: attd a light Otam C'DK- compnmitg the ansino aetd :m\|tmitca ticpti ted tn SL;> ID NO-10!; M a light chain ODR1 comprising the :jro'rnt.» acid v.;qnau:c depicted in Si'.O )D NO:N4,;; hi>hr eham <’ DR) eorapr hnng the amine acid sequence depicted m KEQ ]D NONN eed x hglu chain «.'OR’ comprising the eneee acid set lucnee depicted in SF<) ΙΠ NO. K)'· \'<1) u hebt chain C'DRl o-«mprising she ammo acid sequence depicted in Set.) [D NO' Hid; ;; light chain ( ORd comprising the animo acid sequence depicted ut SRQ I'D NO. 106; and a light chain COR.) comprising the ammo acid Sequence depicted in SEQ ID ΝΟΝΟΝ i;I' a light chain FOR! ..ompr-sktg die ammo acid sequence napieren in SEQ ID NO'Ntq a light chain e DEd com pry ing the amino acid sequence depicusi in SEQ ID NO.HO and a Eight chain (. DR3 comprising die ammo acid sequence dcpicicd in SEQ ID NO; HO; ; vaN a hem chain ODEN comprising fhc amino acid sequence depicted in SCO ID NOQO; a Sight chain CDR 2 comprising the memo acid sequence depicn.O in SEQ ID NO: ί SO; and a Sight Ream CORE eomprNing the amino acid sequence depicted in Sid'd iD MOD 1 K or uiv> a lipid chain CDR! comprising die amino acid sequence depicted in SEQ ID NO.OO: a Sight chain CDi<) comprising die amino acid sequence depicted in SEQ ED NO; 2!: and a light Chain CORN comprising the ammo acid sequence depicted m SEQ !D NO: I So.

In some embodiments, die antibody or antigen-binding fragment thereof compels mg die light chain CDR set αΟ,.ι comprise;·' a heavy chain polypeptide comprising any one of die heavy -.bain CDR ecis m. sot forth in Table N

Id; aofiiièr aypeeh therdNelPsPi'e features ap isoMed amfbedy or Mtigen-binding fragment the roof that comprises a heav y chain CDR set as sot forth in Tabic ) of :TRble S; :Fof OSStople; so yqtftd cmhodiiitehts sit isolated: adtibody or ^iti hhmin eumprlxess heavy eksm pplypepilde comprising; N > a heavy chain CDR I comprising the amino acid mg notice depicted in SEQ ID11 Spa heavy, ehahi CDR1; eompmtng the amine acid segoenco depicted m SEQ ID NO; 144; and a heavy dtam CDSCi comprising the amino acid sequence depicted in SEQ ID NO;11 ?. Cd a heavy chain CDR i comprising the amino acid sequence depicted in SEQ ID 00:20 a heavy chain CDR2 comprising the ammo acid sequence depicted in SEQ ID NO'O?; anti a hea vy chain CDR.) comprising the amino add sequence depicted sn SEQ ID NO:3R; (hi> a heavy chain CDR I comprising she amino acid sequence depicted in SEQ ID NO:;60, a heavy chain CDR2 comprising the amino add sequence depicted hi SEQ ID N O : 161;: tmbdt heavy chain QDR3 comprising the amino acid sequence depicted in SEQ ID NO: 162: fivt a heavy chain E DR I comprising the amino acid sequence- depicted m SEQ ID NO: I HR a heavy

btïi5i OQRa comprising the amino acid «cquenve depicted in SEQ HJ ΝΟ:1ό’Ν and a heavy dom; i' DP) eornprishm the ammo acid .sequence depicted ia SF.Q ;D NO: 1 Xp O} u heavy chain ODR! eomprlshm the amim; acid -cqueuce depicted in SEO !D Né? \76; a homey chain F Diva mmtpromsp the mnhso acid. sequence depicted tn SrQ ίΠ NO. 1 ”” and a heavy chain OORS comprising the ammo aetd sequence depicted in SEQ iD NO i O; m n a heavy chad FDR I comp;mine the ammo aesd sequence dvpkled in SEQ Hè NO \ i >; a heat v chain C DRê comprisem the mnino acid seonenee d^pte-ed in SRQ Hè NO'! ie, and a heavy dtm:t ODRè comptishm the s.mino acid sequv nee depicted ttt SEQ H) NO; i 7; < Qi · o heavy ei non ODR ! comprising the at om o acid m-quoitse depickd itt SEQ ID NO. Uvh a hom. y chain i'DRc emnptDme the amino acid sequence depicted >n SFQ ID NO: S o'; anti ·: hvmy charm E [>Rc comptidne the emitto acid sequence depicted iit SFQ ID NO: hel; tv hit a heavy chum OOR] comprising Uk mmm· acid sequence depicted it; Ui:O H> Oh. j iS s heavy dtnm ODR.? comprising tic: mOn·> ;-.md sequence depicted it: SFQ ID NO: EQ. and s ikavy eh:on ODRê e-'-mpnsmg, the umuw :tcid sequence depicted in SFQ ID NO I I ”, ips t i tk jvy uit tm OOK! comp!cony. tne mmm· a^sd '-eepmnec depxkd in Μ) O .D Ν t > Ν < heavv dsam FOR? coinramu-c tee ammo ecut -equenvc Jepu ted m SEQ ip N; t.pM ,md a he -v v Joe. i DRp t, omm'S-mc. u:e mnum nod monetke Jepsmcd in SRO D NO , i~, m a heavy eham OOR I vViopk.-me the ammo cesd soquenCv depicted m SEQ> ID NO.· D\ a itea\ s Jutm y Ok ' composing Pc «mm ' amd -'v.qisesko depkteo in .v Q ID NO ide: and a houw >.ham v OR Ο otopv:\my 'he ammo aoO <· -quone.' >Rpi- ted m utjO d 5 XO'Ϊ7' i^f > a hemo eheuit. DR} compo-nx ''he attreo «esd sequence depicted in Si O ID NO \ id « heavy chain Γ ORd cont pi tone die an" k> cold setpunkk d^p-evd tn Si ii t Ne: id'.h and a heavy cham {. DR3 ^.ηορποηρ t!>e ammo aod se-.jai.ncs depteted tn St Q ID NO1'! Op <>, h.; a he:o y chain CDR i annpnvntn the a mono acid sequence depicted in 8NQ ;D N(): 131, a heavy chain ODRo οοηοοηοηρ the mnins.· amd sctjaence depicted in She) Γ.) NO; sad. and a I tea oy chain CDR’ comprising ihe nrntim acid scquetice depicted in SOD :D NO: i 3 3; (Cdi > a heavy chain C DR i oontproony she ammo acid sequence depicted in StQ ID NODS; a heavy chmn CDRd LOtopristna site arnitto aekt aeqne;tee depicted in StQ ID NO'dét and a heavy chain ODR3 Cuinpriainp the amino acid seqaenee depicted in SüQ ID ,NO>47; or iN.tvs a heavy- chain ODR I comprising -he atm no acid sequence depicted in SEQ ID NO' 1 .NS; a heavy ebam ODRN comprising the at moo acid sequence depteted io SEQ ID NO: 13ami e. is;/svv chain ODR.I comprising thv amino ne id sequence depictvj in SCO ID NO: Ids. D sonic emR idsmems. the unit body ot antigen binding tmgmcnf ihepnoT comprising Ne hoor \ chain CDR Ssi also comprises u light e has is polypeptide comprising any one ot The light chain DDR sets as sot h>uh is: Table ?,

In ane-thet asper :hc dNeiosme icamrea an isolated -antibody o? antigen·· binding, iragmem thereof comprising a light chain CDR net irotn 1 able 7 am* ns cognate heavy chain C OR sefset forth hi Tabic R. is: .mother aspect, the niseio-uro fotttnrcs an it-.oiated antibody m out igciNNkhng fragment dntrvof comprising a paired light chain ami heavy chain CDR sci its set f»rth in Fable 3 of 1 aide ο K« example. m φ)Μ cRïbbiiMèJbs. air isolaied antibody < Or aisigcn'-biiidïBg: fragment 'dierenfl described herein comprises: ii) a hahr chain CDR i comprising the amino add sequence depicted its SCN ID NO: 140. a heist chain COR.'.' comprising the amino acid sequence depicted its Si::Q !D NO:°6. -.¾ hqju chain CDRc Comprising the amino acid sequence depicted m Si.'.O ID NO: ;4N; u heavy chain CDR· comprising the amino nohi sequence depicted in Si Q SO NO:115;;; hears eham CDi-G comprising the amino no id sequence depicted m SCO ID NO: 144; and a heavy chain CDRd comprising iite ammo aud sequence depicted hi Sh.Q iD NO;I IT; ids a tight chain CDR I comprising the ammo acid sequence depicted in Sh'Q ID NOh-O; a tight chain € DR 3 composing the arm no acid sequence depicted in SLQ ID NON 1. and a light chain {' DR ' eomurismg the amino acid sequence depicted in SI:.'Q ID N 0:.:2: a Heavy chain CDR I comprising the amino acid sequence depicted in SfhQ iD NONh: a heavy chain C'QRa comprising the amino acid ‘-e-quet-ee depicted in ShO iD NO: 6N and a heavy chain CDR?· comnririns: rhe amino acid sequence depicted in $B> ID ΝΟΝΟ, sdii ;i light chain C DR .i comprising die amino acid sequence depicted N SCO ID NORNN a liglii chain CDRÜ comprising the annuo acid sequence depicted in Sr.Q iD NÖN5N a light eham C2DR2 comprising the ammo acid sequence dept·..ted in SCO iD Nip, 15h; « hou\ y chain CDR i comprising she amino acid sequence depicted in StQ ID NO: I 6R, a hcaf· y chum CDR2 esnuprising the arninu acid sequence UcpictcU in SRQ ID Ni): I άI; and a hem·y chain CDR.'* comprising the ammo acid sequence depicted in StQ iD NO:RN: fjv) a hght chart: CDR i O'-mprisiug the moons neid sequence depicted in SF.Q ID NO; RNR a light chain CDR2 conn'rising site ammo acid sequence depicted in SbO ID Nth: :6m a Ugh? chain CDRS comprising site mntno acid sequence depicted in SiND ID NO: 166' a heavy chain CDR i comprising the on one ac id sequence depicted In ShO ID NO; Dm- a -0.0 0 chain CDR3 comprising the amino ,;vki sequence depleted in Shu ID NO; :6¾ and a he.ix χ chain ( DRR s. omprang the ammo acid mCfueoee <L'pk;cd n: SI Q U') Ni); I "0; i \) a light chain tRDR i C"$upright*.· die ammo acid sequence depicted ;n Si.'Q ID 06), I “2. a Hein chain t OR.) emnpili-mg the amino acid sequence depicted in RRO ID NO: I NR a heht chain i OR 3 compO'Iny the ammo acid sequence depicted in RFO 10 NO: l NR a nemo, chant F DR 1 eotiijiiiMne the amino- aesd sequence depicted in 6.Γ.Ο ID NO' 1 "6; a he-ax> chain <T>R.; c-moo: N urn 0-..- amino acid seqnermo JcpieneJ in RRO :D NO: i and a heavy chain h'DRc e-.-mpnMng :Re amino acid seqncneo depicted in RFy :D NO OR; O ii a Fgh; chain CDRl c<-mo: mmo die amino avid sequence depicted in Sidy ID NO'SS a light chain * id,R2 compsmmg Pm amino acid sequence depicted in SLQ ID NORu; a Rent cham t OR a ccrnpriainc the comm acid sequence depicted in RhQ : Li NR1 Ή!; a he-o.\ clash; i DR I uompxmmg the an-Ino ao>d sequence depicted m NLO RJ V 1 Rk a hexvy chOit 6 DR 2 cmnprkiug the amino acid aecincncv depicted I;: SRC* ID N* a 1D): ami a hoax x sham t DR) s emps F inc the 00000 acid sequence di.-ptaied tit Shi) li > NO 121: tx - it a light chain OOk 1 ooenproioo. die amin-t acid ssXjneuee depicted in ShO 10 NO' 105; a Inch* chair; CORD soixipiNing die amnio acid sequence depleted in RRO ID No. 106; a ildn" chain ( DR.2 com;mmum ihc ammo acid sequence depleted in SRy ID NO: I OR; a henvv chain 0' DR i eemprkmg the amino a;. id '.cucncc depicted In RFC* ID \\r 1: fc a heaxj chain CORD vonipnsing the ;nnm<' acid sequence depicted in RFC* ID No. lad; and a henxx chant ( DRR comprising the «nano acid sequence- depicted ;n Se Q ID NO;; ] '·’. -xud: a light cinnn R'S )Rl compr-simi tin. mrmn a·: id sequence depicted in RF.O ID NOyoe a light chain C'Disd evicprlsiog the amino-acid sequence depicted in 5EQ ID N0N5; a light chain ODR) ccropiloin:; the amino ncit! sequence depicted In RRQ ID NONty a iteaxx chain DDR I CannpiDmg the ammo acid sequence depicted in RrO ID NO: I:R, a hoops eh:un 0DR.3 cornmOmo the amine ackl sequence depicted tn SRo ID NO: I IO and a heavy chain OORo comprising the am me- scml sequence de pm:cii ttt SRC ID Ni? I 17; tia : a light chain DDR I comprising the am me acid sequence depicted in SFO ID N ():1.:0, a light chain DDR.) a apprising the amino acid .sequence depicted in SFO H.) NO: 110; a Intht cltah; C DR3 canireNing the atnino acid so>.n.mnce siopicteo in ST*) ID NO:111; a decoy ciiain CDRl omnpnoing the amin-c acid sequence depicted in SFO ID NO. I ?f>; tt heax x chain ii'ORR comprisim.; ihc aintno ac;d oociuenoo depicted in SII.Q ID NO-1and 3 heax x ch:un DDR) cienprisinv the amitm ac 1-..1 sequence depicted m SR óp ID NO: 1RR, <m a iigM chain CDRl compvtsing the aninio a>.id sequence depsNcu in Ski) iD NODE; n l-glu chain ONE 3 con-ps-IOng the amino acid sequence depicied in Shi) ID NO'21: a light chain * ’DR;· comprising, fhe amino acid reen .enne JcpicicJ iu SRQ ID NO. 1 i 3; a heavy chain COR i comprising the amino acid sequence depicted in SR O ID NO,SS; a heavy chain CDR? comprising die amino acnt sequence depleted in SEO ID NQ;46: and a heavy chain CDR 3 comprising fhe ammo acid sequence depleted in SRO ;D NO:4Q ;>.i? a light chain CDR; comprising the ooo no acid sequence depicted in SDQ ID NO:99. a light chain CDR? emu prising fhe amino acid sequence depicted in SDR) ID NO: iEE; a light chain CDR 3 no reprising fhe amino acid sequence duplcied in SDQ ID ΝΟΜΟΙ: a heavy chain CDRI comprising die amino acid sequence depicted in SEQ ID NO; i IR; a heavy chain 0DR3 comprising fhe amino acid sequence depicted in SDQ ID NO: lee; end a heavy chain CDR3 cornyunung the amino aold sequence depicted in SEQ) ID NO:Dm; o;ii> a light chain CDR i cm uprising the amino acid sequence depicted in SDQ SD ΝΌ.95; a tight Noun CDR3 eomprishig the amino acid sequence depicted in SEQ ID N0:9m a light chain C DR 3 comprising fhe ammo acid sequence depicted o; SEQ) ID NON?; n heavy ekm; CDR 1 comprising the amino acid sequence depicted in SEQ) ID NO: 11>; a heavy chain CDR 2 comprising die amino acid sequence depicted in SEQ) ID NO'EDD and a heavy chain CDR 3 comprising the amino acid sequence depleted in SDQ ID NO:; P; t <iiii a light chain CDRI comprising tik amino acid sequence depicted in SEO iD NO: i40, a light chain CDR.: comprising t!;e amino acid sequence depicted in SEQ ID ΝΟΝΟ a light clu.ee 0ΌΕ3 comprising the amino acid sequ-.ncc depicted in SEQ ID NO: 142; a heav> chain CDR i comprising the amino acid sequence depicted in SEQ ID NO.I IN' a heavy chain CDR.? eomprisisig the amino ue;d sequence depicted It; SEQ) ID NO'123; and a heavy chain CDR.3 comprising, the amino vein sequence depicted in SEQ ID NO: 11 ?: {.six t a light chain CDR! e-mp rising the arnnm acid sequence depicted in 8EQ iD NO: 105: a Ugh; chain 0DR2 compnsmg the amino ucn.l sequence depicted in SEQ iD NOMEm, a light chain C'DRe comprising the amino acid sequence depicted in SDQ ID NONE?; a heavy chain CDRI comprising the amino acid sequence depicted in SEQ ID NO: i 15; a heavy chain 0DR2 comprising the amino acid sequence depicted m SEQ ID NO; 125. and a heavy chain CDRo eotqprfsing. ihq amiho acid: seqhence depleted in SEQ ID NQ; 317;. (NR? s 'light chain CD;·: i composing fhe amino acid sequence dcpicfed m SEQ) ID NO:R2; a light chaif; t"DR2 comprising the ammo acid sequence depicted m SEQ) ID N 0:-0, a light chan; ('D;<3 eomprsssog du.· nmuts > acid sequence depicted in SEQ iD NON Oh: a heavy chain CDR i ceuwssnig the am is o ; mud s-queuee depicted in SRQ fD NO. 115: a hemy chain 1' DR.3 Vs>mpfiQtrg the ene:-o acid .sequence depicted in SEQ U') NO: 144; end a heavy cNdn CDR.? comprising the amice sseid sesisu.mee depicted in SEQ ID NON ld: ivCs; a sight chain OOR: comprising the amino at. :<j -K-queuce dep ins cd in SEQ iD MjSO; a light chain CDR? oomprssusg the ammo acid sequence depleted is: SEQ iD NO.DR a light drain ODRd eos-tprising she amines acid sequence sissnicicsi hj SET'1 iD 70):'O; a heat y id's en; t 'DR I eomps'sshrg she amutso red aecpn etsen depicted in SEC1 iO NO:! 15, a heavy chase CDR? comprising she a miss*: ustsd cequessee depicted ie SEQ iO NO: s 2d. and a heavy chase ODRc cos uprising the miiOo acid sequence depicted in SEO :O NO. IP; sew'd) u light chain CDS i curnpt Risp; tbc amino acid sequence defected its SEO ID NO:u? a light chans CDR? comprising the amum aesd sequence depicted in SEQ iD NO:Sé, -.¾ light chain CDR3 emrsptishig tbc amino acid sequence dep; er cd sn SEQ t.D NOOR a heavy drain c'DR: comprising tits;· t;sn Ir: s - acid sequence :.i::p;c;c>i sss SEQ iD NO'; 15; a heavy chans ODRd comprising the amino acid sequesrCu depicted sn SEQ U") NO: 144; and a heavy Chain CidRd comprising the arcs;κ> acid .sequence depicted in SEQ ID NON ! ?, t.svssis u tight chads s'.'DRl comprising the u-n hies acid sequence depicted iss SEQ ID NOON: a sight chads CDR a c<-mprienig the assrino add sequence depicted its SEC1 it) NO: NX a sigh; chans ODRd cm uprising the amino acid sequence depicted iss SEO ID NON 03, a heaqv churn CDR] comprising she amino acid wquenoe depicted in SEQ ID NO: 1;5: a heavy chain ODRa contprsssng She a ns is so acid ‘equctiee depicted in SEQ iD NO; i ?0: and a heavy chain i~ DR?· comprising the ammo acid .sequence depicted in SEQ ID NO:!! 7, tvi.s a light drain ODR! comprising the amino acid sequence siepicsesj lit SEQ iD NO'Ό; a hght chair: CD Ed comprrsrng the ammo acid sequence depicted in SEQ JD NOxri: a lipjsr chain CDEd 'CowprRmg the ammo acid sequence depictedw SEQ ID NO.R~; s heavy ciurin C DR I comprising the amino add sequence depicted m SEQ ID NO: I I 5; a heavy chain CDR2 comprising she ammo acid sequence depicted in SEQ .ID NON.;?: and a heavy chain C DR? comprising the amino acid sequence ska-aered rn SEE) (D NON ! ?; (vss a uglu Drain ('DR1 comprising she assrino aetd sequence depicted in SEQ ID N0.K4; a light chain CDR2 comprising lire amino acid sequence depicted in ΝΕΟ il> NO.N?; a light drain CDRi comprising the amino acid sequence depicted isr SEQ ID N0:i03: a heavy chain C'DR! comprising she amino tied sequence depicted in SEQ ID NO:: 15; a heavy vhaisr COR? eosrsprisnrg the arnisso acid sequence depicted m SEQ 10 NO: 144, ;.uul a heavy chain CD Re comps'ising she anbno acid sequence depicted su SEQ ;l> NO:: i 7; or iss-sa lipid drum CDRi comprising she ammo avid sequence depicted in SEQ ID NO DO; 3 hgfo chain 0DR2 comprising she memo add sequence depicted Is* SEO ID NO:hi: a sight chain DDR.' comprising the asn i ra > acid sequence depicted ie SEQ ID NO; |j ?· a heavy chain

CDRI comprising ihc asnsno add sequence depicted in SEQ ID NO: ids: a heavy chain ODR2 romprornsq she anvi.no acid sequence depicted in SID) jD NO' i .ON end a heavy chant C DR 3 on mens Op the amino an i·: i sequence depicted ist SEQ ] D NO: i.\E in some embodink-ms. an antibody on· muigen-binding Nag mem thereof comprises a paired light chasn E'DR act and heavy chain COR ad a.s set forth in Table N In some ernbodhncim·, ;>e antibody or unmym-bmdmg Ragmen! diet oof cotnpnss-s a paired light uhain CDR net and heat y chain t. DR act as set forth its Ί able 2. For eRspfofoOfle dsscloyurc'leatttfes m antibody eornffoslmg:: fi) s light chain QDRd emnpiising the amino aeid sequence depicted m SEQ ID 00:20,pi; a light chain 0DR2 comprising the urnistu acid sequence depicted in ST-.Q ID NO:2 i: and tihi a. light chain CDRd comprising fee amino aeid sequence depteted sn SEQ ID NO:22. (tv) a heavy chain C DR I comprising the and no acid sequence depicted in SEN ID NO:.:8: ivt a heavy chain CD RE comprising the amino acid sequence depicted irs SEQ ID NO:4t\ ansi (vl) a heavy chain CDRd comprising the ftroino acid sequence depicted in SEQ ID 00:415.

Irs some embodiments. the antibody or aniigon-binding, t'ragtnetu thereof Comprises a light chain: Vifo&amp;hlq fogioq nét: forth in'laffe 2» fohibll light chain: is pahml whh 'My:one of the heavy chafe variable regfeias Set forth' In ’fobfefo for example, the disclosure features an antibody tor an antigen-binding fragment moreol) cemprissng: (s) a light chain yaSshle region having stmamfeq acid sequence comprising the memo acid sequence depicted in SEQ ID N0:d2 and s>) a heavy chain variable region having ass amino acid sequence comprising the amino acid sequence dep t of ed: i n Si df ID N 0 :45. in some embodiments, an antibody or andgosvbmdmg fragment 1 hereof dsserilfod herein ebmprises:: (I) a hènw chain variablerégim tmmêwohfe regiem 1 comprising she amino add sequence depicted in SEQ 10 N0.68 0? SEQ ID NODE; fit) a heavy chafe variable^ regfeu ifesnawsfek tygfon 2 ednfofiSing the asnlsto acid sequence depicted in SEQ ID NO DO or SEQ ID NO: 71: and a heavy chain variable region framework region 3 comprising the amino acid sequence depicted m any one of' Si’0 ID NOs:"!'' to ·Ι in some embodiments, rhc amibsDy or antigen -binding tYagmutn. thereof comprises *· heavy chain. variable region iVinnevror-k region .4 comprising she ammo acid sequence depicted In üi:.Q ID ^0:75, hi some embodiments. die tmlihodv or amigun-binding fragment theruof comprise'· a henry ehasn variable region eompns-ng the amino acid sequence depicted in am one of SbQ ID *40s:?b to 80. The antibody heav\ chain can comprise any of the heave chain CDR sets described herein. The heavy chain \ ariablc region can he. in <*.jmc embodiments. paired with the variable region polypeptide eomprismg the amino acid sequence depleted in Shq !D NO: 16.

In «ome embèdimèpts, dn antibody # aaiigen-binding Ébgmgni Ihereof MMs; to a non-human ('.Ns prolein, !"*>r example, site un-ihody va' antigen-binding Ihtgsnests thereof can bind to mouse €5 a amkor desarginoted mouse Oa protein. In some embodiments. ass isolated antibody or antigen-binding fragment thereof can birul to mouse Cba (and or descry;eased mouse 0 mu and comprise, (if a light chant CDR 1 comprising the amt no acid sequence depte-ed :n SiQ ID NO'.ö-b it ï a Sight chain CDRO comprising the amino acid sequence depicted in beO ID NO. 53; t iit i a fight chain ODRc comprising the umin·; acid sequence depicted in CEO fD NO :5m tiv; a heavy chain CDR I comprising 1 he amino acid sequence depicted its SbQ ID NOm.'h tv ) a heavy chain f"DRe comprising the amino acid sequence depicted in SON ID NO:b.v. and ; 0 a heavy chain CDR3 composing the amino acid sequence depicted in SIi.O ID NO:M ht some embodiments. the ami-monae CSa rmSbody can eotnprise a light chant pob. peptide comprising the ammo acid sequence depicted 1st sifc() ID N0:5v; retd a heavy chant polypeptide comprising the amino acid sequence dcptcled its SEQ ID NO;».,

Irt $<m&amp; cmbMi rtietngs mi isolaied anMbotly or antigesis-biitdtug, fragment; thereof described he sent inhibits the u; enaction between (Da and a ON; ro>.eptor. The 05a receptor can be e.g.. 0500 or 05Lh

Its so sue embodiments. ait isolated antibody or antigen-binding fragment tnetcijf described herein doc:· not snbctanmtily Inhibit complement-mediated heMplygis of fed blood cells: lb \4tm and/orIn vloa, ht some embodiments, an isolated antibody (and accord;itgiv tmy umigeti-bi nditig ftagmcar thereof) Is a moaeelqnai antibody* a lut marnced an tibody;s or a fid iy-human: unbbödy. in some r.-mfco:.iinvnfs. an isobarnd antibody or asngret:-binding fra\isuern thereof described heroin is .wketedlVan-i the group cotvsi-ssms? of », reeonrbinaot antibody a xtsmle sham -subbed;., a dGK-Jt, an innahody, e. cinown/eii or eismtcric anobods, a demursunmed fnmv.n ;nnsm‘h\, an f s frugsnens. an Id ira gsnoist. an rub iragmont. an a s-.b' bo mm. m, and an hab'p unyment in sums. unhodisnenK an ηοίηοο antibody ,u undgmi-binding fragment these-ffskxnbeb he":;·; ts uuibispemik ·.--· :.· , im met. imp jp 'bas. Sim omiboJv nr iVagm·,;n binds m -n kom m ·> ihsiesem epitopes. The two different epitopes can be. >.,p , uuo different epuepm bests the same preien? ;c a . ('em o; sin: xmsh-dj can bind k> a his? ;:p;hme it'ors? t; fits- psklii ;0 g , Oha) u»j a sCCoUn epitope tYotn a mCOiki proto?,», is\ vamc enybodinmns-.-, ;m u Gated andbodv or ant:gen-binding iVapn-e·;· thereof described herein comprises ;·; heter-dogous moiety. foe hesekhoymm nmicn can be e.g . a sugar f os evonpio tin, rnntb-idy or anbgen-hinding intyment there->f can be y:ye-.>m,hned ΐ he hesesoiogotja mmety east be. e.g., a detectable bbe! Mich -a\ hui II"! limited to. a ho< mu tern label a imrunescem in he:, x he;?\ v metal label a radi'/aetive label oran eoeymabe label.

In -emoe etnbodimeitts, an inoiated anb-Cea antibody <>r urtiiymi-btnding iimueia! there·, d'described herein h> mod id eh wisii a moiety ihx; improves the suibbemrossi and or rckobm; of liie anybody m eircutafunt. lor example, the snodibeaiion eun be do Gy labor; or boss bnion. in anoden' embodiment due »n?s-Cxi ;m thong. can contain an altered tost si-ant region that has t educed «Όΐ no'; effector tlsnebon a;- compared die arkxlos' function of ha corresponding unaltered constant region. in some embodiments the asm-Coa antibody contain:- an altered constant region drat has hem eon a bom 0 to about aO;>a id'she effector funetiesi of she unaltered constant region, imenapiary embodiments- of a a eh o:.cre:mc<.befl;cn.'r Uut-..kou antibodies arc desebbed bek'in.

In another aspect, the diseksure features nr: isolated antibody or antigen-hinding fragment thereof that eomsb'locha the binding of xrsy ms·;: o?' tik- tnregoiny «rttbodles,

In f st aitsafber ithpeel tits di selostrsii ibtrktm; a, plarmseetdicai eompohltlou fttereof described herein and dsitsesl. »ö#br excipient. ίη another aspect, the dim lose re features: situ nude ie eed encoding one or mote of any of’the antibodies ot an neen buiding fee erne ate -hereo: described herein; iiit a rectos eosupnststg the suicide acid; ah) no es pressies; vector cosupriditg die nnelek mm, end >·ί ίο, t a evH ceunprisme tik- vector or the expres won veen w. in ane-rho? asp,\r du' >hsohOue tcamssw a method for producing u polypeptide tench ns sns\ of ilk antibodies "ï Simsgen-bommw i'mgmcnts shes coddescribed herein s. The «noth'>d e>nop; ivc:- cduu mg Urn itbresuenboned eel! : comprising du.' ί.·>·. press lost dados') under ©onditioits &amp;ftd: .töMlöW^pfessjêB: itsè cel! of de rmnPeJy o; antigen dmimy fragment encoded by the nucleic add in die a ector. 1 he method eon .sbo include isolated (he antibody or a msgen-binding fragment bom the edi or ftom the sued nun m which the celt is endured.

In ruMhoi aspew, die disclosure features art isolated nucleic acid encoding any ot ilk woum ,wtd *e«.}ueikvs doertbed herein ora polypeptide having an amino add sCijuunC'.. compm inn o; sonswfmg. oi.. any of the amino acid .sevjtsosK'Cs set forth iu-rom fee «mekte .ict-i can I·..- included in a vector, e g;., an expressjon vector, uml/or .005 be ptCsCui u« a sCb

In \et enu.nhc? aspect. the disclosure features a therapeutic kit eomprisistg; sit one or more ol the isolated tunibodies or uniigen-binding tragnients described herdrt le.g., one or more of any of the lunnmnacd antibodies or etsfigen-bitKlifig fragments thereof described herein;; and «ii ? means lor delivery oft he antibody of antigen -binding fragment to u subject. The meem-· cun be suitable for, mg., sobetmme;ms delivery innenendr dvlivory . or uurnaroenlar deli'cry ot rhe am-body or nudge;; binding iragrnen; ’hereof to rhe -mbieu. The means can be,. c,g,, a syringe, a double· barreled syringe. or two separate syringes incorporated for use in ?uhn luistering a therapeutic ant need or <mo pen-binding fragment thereof, while drawing off knee fluid is-.g., for analysis; in a push-pull fashion, in some embodiments, the means is for ocular delivery arid comprises a oems-sclera I patel; or a contact lens, each of w hid; comprises the nmiboby or antigen-biiKllng fragment thereof. In some embodiments, the means is stumble for imrapulmonary del-very. For mmmple, -he means can be an inhaici or a nebwisee In some embodiments dm moms? ts a pre-fillcd syringe such as a pen device. The pte-ftUed syringe can contain, e.g,, at least one pharmacetuieul mb; dosage form oi one or more of the antibodies or antigen-binding fra gone sits thereof provided- herein. in some embodinvnis. dk- therapeutic kits described heroin cat- contain at least one additional acbv e agcul fOr use m treating a complement associated disorder In s subject The addiftomtd active ugem can be. 0.¾ . an> of the additional agens-dc^ciilvd herein

In yet apoiker aspect., the élsclosiïr© feann-cs a method lor ucatm^ or psmonnng aeotnpien,em'.js,v.\Kned disorder Che method meindc' athnupstexug to .: human m need lueteot a th. rapeon·.. .mub.nh o, antigen binding loo-room there·>Γ d’-v nh. J h. ree' m an amount sufficient to own a e'imph-mcre-nssoeliKed diender .dibebng die hut'.,o' The method e-m tls·-· hkltide tdenefv mg 'die subare; -w hie, mg a v o sure o n ie s o - e \ -,. ό i a t e< 1 elsosdu, The , ompiesuesu associated disotder can he, e £,. a eofoptenviU-ess'ieoneii mfhninn.notv plsordos .avowal hemolytic niemk wndiome ay.· minted oucubii dogenoTam-u, rheumamui artinitts. sepsis, or asiiiphosphohrnd svnmome hi some stniwhnkins, die e o: i s r Im n e no a ss oe 1 a e d dia<<uk; is ., eoinpieniann.·.:'soeiatee pucmmay d-soider her e\ample, the ..osuplemem-,sM.viatesi pueumuuy 0;sorJei o in i ; e g , asthma or chrome obsouenve polmosoip, disc ι;Ό. Other complement ax-oeniLd disorders amenable to treairoent m prevention es set hath to die met hoe me de.O mod hofsits Ine rmnie of admmistianon, o Inch Can wP, depeedwg on die ο,ρο of emupkiniM assoc mud dkorde; to be Kerned. 0..,0 be. e y mnes a nous tummisn.itset;, mb tpu imonar) administration. Intraocular lull:" nisirution s.hvuuukotss ownau-ttnoon ot siiinuutk'ihas administration. in some mohodhoeuts, dte aa-d','-dv O; amigon-biadisy fragment. thereof is administered to the human iti an amount and wit!» s.t frequency sufficient to runmuihi a reduced level of sysiotnic C5a «etiv by lor the dusatki·; oidhe ttetinneat. In some embodiments die methods can mo lade idler the aurninisiormg, monitoring the human för au. 'MptméfOimï -¾. öaé or more symp toms of the oonqslemenyaysodiaieil disorder;. in some embodiments, the methods can include administering a- site human One Or more additional therapeutic agents. in vet another aspect, the disclosure features an article of manufacture, which comprises: (i) a ··»#*.%%Μ·aast (ti| a composition oompfisksg on antibody or unkgen-binding fragment -hereof described iicrdn. 'fbe label can indicate dun dte comp·. :sirioii is to be administered to a human having, suspected of has mg, or at risk fer developing, a comploineoi-mssoenttod dtsertey The article of ·ngmufaeturs esc also include one or more adeknona! active agents. Λ.·» n-ed tbronynout tlic present di-closeie, ;Ho term A; nobody" rot ms ;·.* a wh<<ie ;>r sïiï-ii.ï antibody ->o y, lyy'sl lgG lo. \ :yD. or iyr t mokenlc that ss ^oncratèè b\ any >ffK 'if-, stifict) oi nktbods th.it me kstown m the m* and dtaCHbed herein. 1 he ie;m 'UnPlv'dy ' include- a pOveleoal anihedy..;i monoclonal .inttnodx . a OH; re en orcinmenc antibody. a nmnanmed antibody, a demironnmed harnas antin'>d\, and a tally human «nilhedv 'Π-e antibody mat be made m et domed froth >ave '-fa '.assoc, ol species. o y , teen cnai snob as in.; mans noo-iumtni pnome- se.g . monkey.·» baboons, o; omrnnan-ves' horse"' emtio pigs. -neep, yon-s, dogs, cats:, rabbits, yumen pays, yes'bils, he trades s. iet-, ,nnj once 'The antibody can be a purified Or a meohdjlsaot antibody,

As used he sent. the tes'sn "antibody fYaernem,.' GmbganbisKshsy iragntcnC’ t>r yimilar terms refer: fma ir agsbeot of ao:a»tibody that retains Che ability to bind lo an aiitigon (e.g., art epitope present isvOSib but not ih the a|pba oltai» of uodeayetl, stati ve C5 protein·, i:.:.y. a single chain antibody tscFv O an Fd fragment an Fab fraysnem. an lab' daymen·. or an Half ),· fragment. ArisoF'·. Is a sit-yic yniypcphde chain that includes bah the heavy and light chain variable regions et'the a nobody fro so which the seFv is derived, lit addition, diabodlos {Poliak ; 1004} ,Voo< env 2( 1 2?~. 112 M 12m Bhifaoh :Cth|. Clf Immunol Aimfmis· p III 41; I? ?- I hfk the bbolofurcs of both of which are incorporated herein by reference in their entirely b srtimbodies. triabodies I Solis!'sni.'i.'ghe et al. t2000} BX/C Bkn^htuB oGOy domain antibodies ialso known as shcsvy chain· Holt et sf. f 2001 it if IGfufy ffetóefeiof 21; 1 U:4S4-4vO;·; and jtmabodk's (Huston ei ai. {200i t H'im ArOhoaii·'; 10(3-41:127-1 -12; Wheeler -J ai, (dia·.)' .Uol Γόα' bGUGohkk'b; Stocks (20()41 />mo /Xvov 'fmmw m22) '->o0'!h.', rhe ihseh-ines oi e-a.h of which mv ineorp<msk\i Ικη.ν, by t ok voce ih their entirety] ate incioded to thedsInitsoB: of antibody fragments and can be incorporated inn- the eon-positions. and ased in -he methods. dsmer-bed herein.

Unless otherwise donned. ail Ice ins scat and sewmibc terms nseu hosvm have tlte oartre theahlagsas eoutmouly nndhrstood by one s>f ordinary skill In the art: to which rhis ilisel'-aare ps-rtains. In emte of conflict, the present document, indndnty deitriiitmts. w iil eomroi. i'r>.-fcssed sneUtodn and mnk'riciv are desenbetl below. although rneb'mds and srsaterntis s-intlur >jr aysivaiem u.) -hose described beam e;m also be: used in the pittedde or tcsiiiig of Ihe presently discloseb methods: ancl: cornpo<ns>.-ii)s. AÜ pub; lent ion;-, paten; applications, patents, end -sher references mem Gated herein arc mcorpursied by tetefeuce h; their etttlrety.

Other I curare.; and «e vtniiage-, of the present disciesua'. en:;., methods for treating o ennpknnestu associated duvaafers ut a .subject a if ! be apparent front the ;oH<o<. in a description, the examples, end from -'he claims. brief IX-seriptton of the Drawings Ηί». 1 is « Wm dsags>un depicting the decree ed'ore; kip of!he epitopes in human ( Kneed by a select set ofirwriite ami-human id>a a mi bodies fan!0f Mb. 5 :ι η I dO k ί [-., > am{;· ihM n. fan! 'Nd I b and. '-ar·, f ~8 Mb

Fig. 2 is a hue graph depleting the antagonism o; \'5a ntedu-untg .signaling d; ru"o using a neutrophil aeiix atmo a-uu·.. The V -axis wpreseuts the upfiea) density (fïD- rsHustuemesH ot a chrom- we me substrate as a function of nerdopuroxidase release by freshly isolated human nemrophd·;. 1 he X -exa- represents rhe e on eettt rut ton - -T .usi =f.-< -d\ incubated o 1th ύκ· cell. 1 lie human wed antibodies tested - RNj.'-'i". BXj.mT BN.; d f B\.t V"y. Rm ΠΚ;. i.F\JhS.g ami a hmnanir.ed anit-CS antibody an. identified tit the inset h,need.

Fig. 3 lx a hue graph deyilmwg the cl lee; of several therapeutic an ripen ter on jbidf intlattspjMph ih a tabbse «tpifei; of dtensMfoid; dtlhnfas: the duck»;.--: a of the ; a dimly im'lamcd Knee jeinl in millimeters. The X-axis represents the days dftet' diseh&amp;e onset The theiipeptie ehtiftodies tested ·- Sami fSMIE C$ biobse arm-mouse C5a antibody} and a control muTody with the same be reek.at us the a»n-C5a orhibody - aw Ido a dried in the inset dm end.

Fig. 4 is a itite graph depict trig the eft eel of several dternpeulic unbhodtes on .évd?i1|,-f||$^è:.4:#éiliy id a mpesd tppde! of#e«maipi«! trimis, The represents the aril and-; index The X-axis represents the Jays after disease onset. The therapeutic antibodies tested ~ :ταηΙν,.ΝΜΕ <a nunt-w rmii-rnouso C5a antibody) and a control antibody with the same Fc region as the aosi-i'.'Sa antibody arc identified in the inset legend»

Fig. 5 sets forth a scries ·.T;nuvunnwd heat s Τη am \ana Pie region se.incnecs, In order from upp».revst w lowermost tin.· hr m y chain tan.dole region -4’tie: BN;' -is huinanvcU xtidi-s'5a antibody iSFrp ID Nr) Ti dtv kwo > chum \ .limbic region of the t’VXj'dó hiununt vd anti t Tn antibody t Set) if) NO ^1. the hear y chan; variable regio» eftfts H\e t"' humanised mm I'd a andb-od'» -,Sb'v ID MV'-'s* dn: h-owy eh we wninbie region of the B\ Ted hum a nb ed anti r c-u antibody <, S b.O ID NO 'x,\ and the heavy eh;uo tarinble tegten of d>e BNJo50 uumwu'cJ unii-tpe antibody tSI g) ID NO'Nfj ï. Th;.- skilled kx-lbaii v, ill upprer kde the delineation bos o o-m heasv chain fYamcrvvork regions I. 2. 3. ;m<H and Sire heavy chairs f'DRx !· 2, and 2. Such helme/Uw-o- as defined by kabm c; al u^tra) are xhovvsi in the figure. "RC FRT' re: er-· :o heavy dia in variable region Inuoew orh regiem I, 'TIC FR 2" rei er-· :o heavy oi?mn vat om Ie -eg me fkrm.eowrk re eren wk s-kj ' vercrs -o heavy dra er vanahlo meson harem ek. wci-m ' om: -;;(' p kb' retos r<:> ku-avv e kun v unable :s n wsm-g seyon -1 'n-k' ί l>k teksv te he o.v e; am smukte "ogion >vt'spsomem,ou' dek"m?tkrk- teeion-ó'lik:- s Mk s PR v mte·* te ho.oy churn varmbk κmon ΤΤ>Τ~y asrj 'Tb COk.r'refess Ue;n v eh-nn vutubk region e Dr<è

Pig. U i'· a line emrh demetmg the tvkvt-uge ei eucsikcin» tseustvelnk m ih«e bleed el mue iuM-mum 'dsm'-womtcn ol '-λ'\ι ·ο tbo Ι)'ι> e dn She S -adv tkytropht eoarts are e\r<i*>-;</J ;· a petvemutk' ei' baseline," e Inch :s she stessnepiisi cetisis at time 0 {*.-r 1 kk;>;> tsexkophiks. The Xmxis represents time in mintstt. - Motsxe cohorts were intravenously üd-siinkren/d n e-nsrrot antibody [asm-ashhrax ptoioence sniiigpti 63, ϊ.^3;Ί/βί4 RötYpo] .pèèirtr^» five inïcg) öi The.sfikkhtmism CS&amp;· antibody Blxjèk' at one of tho folios ing dones: 24 mg-kg -five mkx"; 12. sou kt> itUe mice); 6 I&amp;ter v?em administrated hC5a.

See nxiunple ΐγ $k mice, "sham," were not administered ho titan (2 5 a

Fig, ? :s a bar gn-.ph def'ktmg the sswelopcto.xujase the!PC'; lord ns she plasma es max bob-re and Phi-or ley aunoni-OatKW o*'human C5a to Ilse mice. The Y-axis represents the '„oneen?-vriest mg min of MPO in oi-new plass-ms. The X-axis s'--presents :m:> In mmm-n Mouse cohort.- ο -,-re ts tr.n crsniudy administered a control nmiboek [tn-ti-eikhtnx. ptokctke antigen ¢-.2 ledTl d-1 isthypej (“control", eight mice;-et die ami'human < '>« antibody IVxJ *x3 at one of five following doses: 24 mgrkgtfsvc idled)!.t£.fcig/Hg·'(five# 10¾¾.(live·rsicey astd J m®ifeg (five: mice) and then 'lais$"sham,5' were- not a-lniiitistcreri human (Ida.

Fig, 8 ts a iiste graph, denicStsty ‘die change nt Imsm?t5 C'5a level in plasma of msec (administered human ("5:-.0 m tlte presone·.. --r absence oi dii-en.ot ee-n'-entmtloiis efau -aufi-hCba mol body (BkhlsX 3} The Y-axis rcprc'-cnts the eon^entratkm > of hC'5;·; in mouse pktssrta. The Y-axis represents time in minutes. Mouse eohorw. woro * co.n.tol atiabody [aktbatiihmx. proteetive aiiigctt 63. IgG'.T.G-l isotypej F'eontrof'. six mice: or the ami -human C5a sssubvxty BNJ263 at one o: the 1btk>v\tna doses; 2.4 mtyky tthree smec): 12 mg-;kg;three mice:. 6 rou/kg ills τα· mice): see 3 mg.'kg f three rnirei and then bier wen.* idrninis-rated hCêa. Four mice, Wham,"' were me administered human C5&amp;.

Fig. 9 K « line graph depleting the competition for binding u> human C?a ;n wiïth 2SÖ pM MTiïfhehiUpek|sekd amnCAa :W·^·wii|i 1 nM. biotinylated Itt'.Sa. along with various concentrations te.ir., 40(), 133. 4-1.4, 14.3, 4¾ L| and: 0.5 hM| of on e of the folliewing; (a}4mthan: Ops desat-g: protein in pbösphatmliMlïéréd saline, (fel liuffiair:pla$ffia, (e:)::éyödnmlgi.ïS niMaiprd plasitta. Id) Shfh/Cr (Mouse) plasma, Mhhtespeei to the plasms: compotienih (hfe (ofe Id y. and: lei the eonceittradda refes # file approd mate And ednoeeltaiioa óf C5 duigen in the htcobaiion mMtpy, The %®®$·xepresenls; arbitrary ilam'Cseenee unie: as a function of the amount of rmhernumdabded anti-CSa antibody detected. The X-axis represents concentration (riMl of the antigen ebmpefeisit-.

Fig. 10 in a line· graph depicting the ctrect of .several complement inhibitory proteins on the alternative pathway |ΑΓ') ofeomplemem. The Y axis represents the percentage of AP complement activity as cornparal to baseline tBL; the level of sethdiy in the absence of a tpmpJhmerst InMfeitork Tile X-exb Tdpmseofe:1he ooiietsimlioii. ei a given eorsipletnept inhibitor (oMi). The efficis of die smiMCJta antibody:, ENjiiBf, aipng with an anlhph infeifeody on AP activity were each Ovninaiod.

Hg.n lx a hoc graph depicting tin.. effect of several complement inhibitory premini on the eU-wieal pathway f(.' Ï1'· of complement. The Y-axis represents the pvreemage of i.T complement activity as cm owned to hasdine sBi..; the level of iiCti-'· tty in the absence of u compfe-mcn? inhibitor). T he X -axis represents the concentration of a given complement inhibitor ύοΥΙ ·. The ·..-fleets of the ;uui-hOa unilfeodv:. BAd3fe3, along with tm aPifeCS aititfeody oti CP setivify were each, evaluated.

Figs, 12A, 120, I2C, and 12D ere usenes of chromatographs depicting the retention titties ol the aitn-C5a antibody iBN.BHT) astt! atiastl-lilf5 sntlbody hi tile |5r&amp;t^tde.;pr:lttehee of hi.' 3 protein for all of the panels, tim Xopfes: represesiis retention time in minutes and the Y-axis represents the absorbance units at 21-4 nm svavelength. in each pend, the in laved subpanel depicts an enhanced view of the ';ftóiwé4.;|5délta;-

Fig 1 ?.A depicts The refetnmo time for BNJ383 in the absence of h€5 pro.tcm.

Fig. 12B depicts the reieotlors time tor the anti-Cf antibody in the apppec of FC'f protclp, I' ,g t X ο/'ρ;·, 't\ she wntum fim. Sdr B\ 3h 4 ,·; the •'Vser.e·' >f ht ' i.2 ï -t.'U snoku' evess -n >t oves BVA'! from units t<· wk, the .'nametasce pe.-.-iS lyiV'di! «go usn.ompk\ed B\J ’«.>r hi -3; A* tt\J A;; wsu! one a'mgcn-Xmdtng s-v i .<>ι·ν fu -nvic oeo id v and te' a tv., .or ttaeti-.n 0"ho;--''CiU w >th dad γ οηρ,οΌχ · >1 BXjkst w:h u:'.el.\o _d ttC5 I V i 2D eepteto dk κίον den lone k\ the and t ' asmboek m we pro,-.nee oi a ' eo,. molm oousu .o'hi *' \ rem right to k'fk ho enussvmkJ pucks rcpscsetsf «as; oe<<v'pk'ed ant? * 5 <χηπΙ>·>α> w ΙκΆΒ'νΙκ' mit C5 omihdOv Va'' <»nce 'tigest-moding sr,c boussd to ,;;vL\oed id 5. and {«.' tiv atr.H amihtsh -t-iv d to tv.- uneLvoed v's m<-iodides Hg, 13 a one grapt' o.e-v t "g f'-e bmdtstg oi :iv antt ·Γ> i .untbobv B\ f Is 5 n> h«f5a Jcvu^ η· Ρκ ί'Κ,.'-οΟν e: h< '5 using an FI ISA i v. X -cos tepresent? tee eonee:oi_si..>u «no u-I ' o: fhe to Somov. Foe V-vs"- septesestk she uptMvd Jeststt'. at Ί^> um '.> ..o ckogth.

Fit», 14 ;s ,i scores piet d'pOong she eeoeetstraoun of 1u\ Cv tfé1 ekvatg ps m ihe oVum ai i. vno m> ces stsaeupaes a huseuok ei me ffa-nv con. emtarsost of .mtke ee a -phoov t ó\ i At; t h.„- \ aw dep'et' tin.' cetteem obtest *pg ml : .'ft ''a ( a dern*g dcfecv \ sn pi "sT«t. . j.rop Vs trom . \ sv-m-dgu\ mac Όίκ". at tune po'-'hN sanem>a *i.>m i da·, ''o doos k-ihming a Mngie Jo-e imr e. enoto ad!5''ne'icunm .'FBM'i^> .-a i ->tg Lg. id r-g kg Hxo sag kg„ f "Ό eg k.g os Ao !0g kg bed) .vtdghi ...>f titc antina In. i lie .X~axk represents the eoncetttnttion of the. iXNjjHa fug ml) in each· of the samples.

Fig. 15Λ ts a seal ter pirn depicting dse percentage of hcmokitc aetk tty in senarn sarnpics relative to baseline values tV-sxisi initiated via the classics! pathway ;.i\ a ft! net ton of she conoco tratifit) of ;nsf;-C5n antib-dy (BN) 3 83) in trculapoo tX-nxts).

Fig, 13B is >t scatter pha depleting the pereernage of terminal cornpkmem e«gmp!ox fortaafipn fniliated Xia the classtcs! pathway tn sontm:samples meakufod by a OifPeo assay reist ive So baseline values (Y-axis - as a fnneti-ot) of the concetv.iarion ofaisti-Cöa anttboOs (BNJ383' its cireulatli.>n ί X n>i·. ?. hSï isw i, n semiet plot Ju; het me the percentage oi terminal '..omploment eömples Ibrmaih« inn luted v o- die ciUsMeul pathway m νομοί sample.- measured ly> a OCP sssay reis the to baschne 'Utues vY->im\) a- a fuueiioa et'the eoits'emtaliou o; aWtlWdSa arhiboby (HlSIj38:3] is ebx;uladon iX-asis;.

Thf psXyöaf diSclosars f?-by|de$'.«n((bodies. md mb igeo4tmdl% imgftWöbs thereof that bind t:.i irer.· C5a to o., tree human Cüa-, compositions containing tbc amibodie·: or theh irug'mems. arid methods for using any of the foregoing to treat nr present co o eb e n um o a- - oc i u te d disoixlera sneb au, bui urn hmHcd b>, aHl.'S. macular degeneration te.g , AMDh RA. -opsis, amiphospholipki syndrome. burr; te.g.. \excre burn],. ipOéiljpast^ne^N^mdibiti^ ^us· ösphrids, or a eOibpleotoabaasbeialod pulmonary disorder on dr as aohmu or ehsvme obsiruchve pulmonary disease iC'OPDi. The chsd'>:-«n.· dsn provides <mii-T'5a nun bod c\ tarui impmeud thereof) that arc c roNU-rcac-bc (-'.tween hvc Cca from human and free 05?. from a rum-human mammalian spec. ics such as a non -human primate (c.g.. Csmommgu- mucauue or slwsus osaoaquey iWIble Is hg way isbebded to lie iimhirtg, exergpiary aarbBodlös f asti umigeu-bmdiuy boomem- c compositions te.g., pharmaceutical composition'· ami ibrniulaliohsb and methods fra- usmg ffce-%£f .tiy^éïöw·φ|<| exemphCkd in the worlastg Examples.

The disdosura provides antibodies dm bind d complement component ('on.

As: dtscnaseii:.above., the pmAtm of C5» s 10:¾ amino acid residue jwepudor prgtöh, ;s processed by -» series of proteolytic cleavage events The first I s peptides (numbered 1K -o -d ) constitute a signal peptide duo is cleaved bom the precmvvr protein. The remaining U-SS amino acid oreturn is eioavod in two places to fem) die alpha and beta churns The fust ebon age event occurs between amino add tcsldncs o55 and shod, flic second cleavage occurs between amino acid residues (>ov and bbO, The two cleavage events res ah m the ibrmtnion oi three distinct polypeptide bygmemw |i) a fegment gompfiMbg ammo aesdh i ® 6¾ which A mfemd to as the beta, eltaiii; fit) a Imgiuefii: comprising acids bbb to 1()38:, whleh is refétycd to as: the alpha cham, anti i'-n; a tcirapeptidc fragment con sis;; no ·υ( urnn··; auds (";5b t·.· bps).

The alpha chain asd the bets chain poiypcpdde iraemems are connected to each other via disulfide bond and constitute the mature 05 protein. The OP or ΛΡ 05 convertase ach'-sves mature 05 by desvmy she alpha chain beacon residue? 733 and "34, which result- m dtc liberation of C5;t fragment mtntno acids <k>0 ns ? ' 0 T he remaining portion of tranote 05 is fray mem Cf>b. e, hich contains she residues 73-1 to K‘55 et -he alpha elnust disulfide bonded 10 she beta chain, />; e<Vo, (..'5a is rapidly ntctaboEUred by a serum enzyme. eattroxyirepudase B. to a "3 ammo acid form termed "05a. detsarg." s\hk.h has lost the carboxyternut-ai :wgmme residue. Accordingly, in some embodiments, an antibody that bind? to free 05a also binds to desarghaued C5a. in some embodiment?, an antibody that bind? U.s Chns does not bind to desarginated ('5a in some embodiment fhe ami-C:'·* mtnbody bind? to a ncoephope prerein m C5a, i.e., an epitope dun becomes exposed uposr the liberation of C5a from lire alpha chain hoy mem of ntaSnte C5. That is. sn some embodiments, an a;;?i-05u audio eh described herein bnuk- to C5a and-vr 05a desarg but ttof to mtciew, ed, mu he fully-· ft skied · ¢.-5::

As described above, in some embodiment?, the aml-05a antibody or antigenbinding 'fragment "hereof can btnd to a subpoptiluiion of uncleavedL processed C5 te.g., plasma 05} consittmutg ices than 10 te.g.. less slum 0,5. O 5.5. 8, 7 5, ", o.5, 5, 5.5. 5. 4.5. 4, 35. 3. 5.5. 5. 1.5., 1.0,5. 0.4. ((3.. 0.3. or less than 0,1 s % of the toud pmmLokm of full length ('5 h; >·, sample te.g , rt blood or plasma sample or a sample . é4^Ci$|h.g;· fü-II Mögfti 0¾ sybiciisubpdphlddbd ImM whole, , bifid pail, dcnaluted such that ;m -nhensise oeehided (35» neoepilope Is exposed. Thus, an anfl-5 us antibody or amken-binding fragment the mol described herein can, in some . embodimen ts, bind to Ifee 05», but n ot to €f of the ttr.sgraster.n'ncl^M, :nsdy e 05 population. In some embodiments the pari tally or fully denatured subpopthafion is inueihe or hut reduced act:' dy e.w. less than 40. 55, his, 75. 70, 64. 60, 55, 50, 45. 40. 35. 30. 55, 20. I 5 Of >i!:, eh she activity of luliy-Ametlanal, full-length CS protein) in any number of suitable assays use in i So-r testing 05 aetieity. e.g., a tanolylfc assay or a CM50e(| ahsisy, -Smia%lbdij9tfa.od$.tsstbtg the sbbvity of ire minor snbpopuhdion to which an anii-Opa antibody described herein may. in some .embodiments, bind art? known m the att astdedoyefibod herein. in some mi-bodm-euis. die ;.;n1i-C5a snithixly binds m a mammalian ie,g., human} F5u p totem. For esamplc, the anti-Oda nrmhody can hitui ?o a hmwm Cm; protests having lire followhe inn mo acid swquoitcc: T L QK Ks FF i AA K'Y SCH FV'V KKCC'V DGAC'VNN D FTC F Q R ΑΛ R i SLGPRC! Κ.Λ FT F SfC VYASQLiCANiSHNDMQi OR s$KQ ID NO: 1). In some embodiment!?, an anU-C5a antibody aw hied to a uesargmatul human 05a protesss [saving the following swsm<> acid wane owe

Ti..QK‘ K i i:K ΪΛΛ KY K H S W h'KTCYDOA OV N N DF! CEQR A Λ R; SLGRRCi N A FTF. (X'VVAS'hLRANiSHKDMOLG (SFQ ID NOO: >. An »n!i-C5a ;ms)bod > described herein cim bind to bod; listl-length human Che and dewargissuted hi;man 05a. in some embodiments, the antibody can bind in human 05a ai an epitope within w overlapping witst a structural flagmens oi the protein having the amino acid seouunee; 1 l.OKlw ΓΗΙΛΛΚΥΚ t »f.D :P 00:5 i. HSWKKt. i. YDGAi. lSiOO> iD N(V,4r VON OF sSEQ ID NOOh TCFORAAR <SFQ iD N0:6V ICr.C ;SFo ID COwy RROlKAi Tf-OOVV.-VSOi-RANlÏï (SKQ ID NOrfci: HCDViQf Oi ,SF.Q ;D C0:9s, or HKDMQLOR -SFC Π) NO:iOi. bee. c.g.. Cook ct ai. ; A):0; Act'·.; C ;v« Poo. IRQ-1^7. hi ;.ome embodiments the embody can bind so 05a at an epitope within or overlapping wtr.h the ammo acid scijuusses o; a peptide t rap mem of'Cha c<-toprising -n icmt two of she paired cysteine residues. For csample. an asni-Cèa mtnb'.'dy can bed so fragment composing, or consisting ;;f. Sim as nine» acid seqtseuce' 00 VDGAC'Y ANDCTC sSf.0 iD ΝΟΊ i OVDO.VOV^NDP rCbDRAARlSI GFRC IK AFTFC ;SH) ID AO:: A mid OfrORAARISlAiPRCiKAFIHOC r'sfrO ID NO;Lit. wherein. in each of'the peptide fragment'··, the first and iinal cysteine residues asc paired by disulfide bonds. In some embodiments ;m wPi-Cha antibody described Iserein can bind to a human C5a protests at f;n epitope w ishm, or overlapping w irh. the mntuo acid sequence; YOCACVNNDKTC EQRAAR <SF.Q ID NO; Ml or CYlXiAOVNNDKrCFORAA sSF.O ΙΠ N0.15) in some ernhodsruesUs. an antibody can bind to a human Cha proiein or fragment thereof comaming an amino acid soquonee that contants. m consists >?; at lea^t Aurr ;,c g at Past sent, sir e \n sew.;;, eight, tunc 10, Si. Sg, i A 14, i 5. Ιό, οι i 7 o? more) consecutive arniru; acids hep clou in asiy one of SFQ ID NOsts to i 5 In some embodiment's, an mni-OCi antsbods described horesn bmds to u term;ry epitope comprising two or more se.g., ;u least ms., three, or Fur) dike ónd nbohy pèpEde fdglbuy of ESN phefem, eVgo $*ffl or moré dlssèöilsi;Pötis ¢5¾ peptide region:: joined together by way <4 a disutbde linkage.

Medmd- for idesniiyhig die epitope -o which a particular amihodv ;e g„ un ami-Cw, umlbodyt bind? are uiso known in she an. For example, die binding epitope e;;dlie 05a ; or desmgosaied CDs; tv winch an am: (?>;. ;-rcdbvdy bind? can bh ideptlflsdlby mfihody to severe! Eve,, six, seven. eight, nine. 10, 15. 2i). or 50 or scorei overlapping peptide fragments of a complement eotrtponcttt 05&amp; protein te.g . avveral overlapping fragments of a human ('5a protein haxbtg the ammo aetd sequence depicted in $1:.() ID NO.'I or ShQ ID ME1)..: Ehbh 0ng peg fidex Igth&amp;mhburgt fe ay® iqug address: mi a volid support, e.e., separate wed:' of a multi-well assay plate. Next, die art?t-('5a antibody lx interrogated by contacting it to each of the peptides in the swsay plate for at) amount of time and under condition? that allow for the antibody to bind to Ha epitope Unbound ond-Ofa antibody is removed by wanhinp each of the wolK. Next. detec tab! y · labeled as-eondury antibody that binds to the anM-Cpu antibody, if present in a welt of die plate, is contacted t<- each of the wells, and unbound secondary andbody ls removed by welling sidpy, Tire preyesee pr amoitrd M (heddidgfhble signal produced by the delectably-labeled secondary antibody in a we It is an Ind'ousou shat the ami -C5si funio; Uv binds to the particular peptide fragment gsyobiated: with the well, iSfeey e,g,, IImIow and itaae fybprxi)» .JL &amp; t# {ώφτφ>. > and U S. Patent Application Publication No. .M0AO15M wy the disdosute of which is Incorporated by reference in its entirety. A panicolur epitope to which an antibody bittds cast also be identified using BlAcoie chromatographic techniques i w-v. e.g., Fkarmaeht Bl Arechuoiogy Handbook, 'hiphope Mapping," Section 0.3.1) (May I ‘-b-H and iotoemt in some embodiments, an nnit-C5a antibody described herein contains a specille set ofligh; chain complcmcntitnty determining regions (CDRs) and or a specific set of heavy chain CDRs. Fo- example, m some embodiments an anii-ii 5a antibody or asUsgesvbsnbdg i'ragvtieni thereof described herein ean comprise a tight chain ODR w·; obtained from a tight chain polypeptide comprising the amino acid sequence depicted in any one of SB) ID NOs: Il). .0,«»? 42. in some embodiments, an antbiBa aMibody.of anllges'binding f Mgmeni Éem>fd^<^b^hh«rpti'eaa eotnpriso a heavy chain (' DR set obtained from a heavy chain polypeptide comprising the amino acid sequence depicted in SEQ ID NO:22 or 45. h.sernpbry light chain and hr-«v'Y chaw c'i}R \oo obtained trom ;he aforementioned light chain variable regions and heavy-cttam variable regum®» arc described bekm hi more du uil tseo Tabic 1 j

The exact bmmdanes of b DRs. and framework' regions, ba', e heen defmed difdretuK ace;>rding te dit torent ou-thr-ds irs sotiu.· unbodimeuts, the positions o; the QI)R£whhiii &amp; light pr- heavy dhsÉt fsifabte dötpdh cm bh te dctïned by Kahut et ai. h Db-M “Sv^uences ot fbc-tems oi immunological m te es ld' NIH Pnbbeail'fü Ne ;>l· ^342, L.S. Department oi i hrahh and Human Services. Bcd-teda. MD1. In «och eases the CURs eau be referred ίο a.s "'dabei C'DUs" ie.η.. "Rabat l.b'DRe'' or "Rabat H€ l)R ΓΊ. In :;<>mc embodimems die positions -T the ('DRs of a light or heavy chain variable rogl-w eau he a·.- defined by Ömdna et al t i VisVi NtfiViit id.V.b’NN'dda. Aoeordtugfy the-w regions eau he reu reed ïo as 'ChoShin CDRs' i e.e . "{ hetina LODK2" oï T hothia slC'DRe'd in seme embodi merits,, tie positions of file ÓDRs of fe light aad heavy cMlrt variable rsgibtM cao be n\ defined by « Rabat-? 'hoSA; combined definition. in sneb embodimoma these regions e;m !-;.- refestvd te &amp;' 'Vembmod K:tbat--Ohoiht.-i OM<s5' hi some emhedimcnis. dis positions < - fthe CDKs and w fminev,otk rcgf-n-i within a tight ot heavy eh;u?; variable dm won can he as de thud by Honnegeer and; FiüeUhun bcOO!: J \Joi Bii'l 3(1 b be "'-(ΑΌ hiciiditeation of die CDRs w Ithin a Ugln vhuin or hero y chain variable region using (be af neme mi·-ned demmmns -s oei: Known in the an oi ahtlïödy engmeefing, .Far example, Thomas er ah f 19961 4/m' ImmuwöÏ dlk.l.êd.LNfl 3i';9-1 TO} I exemplifies the idcmifcmhia oflignt chain and heavy chain CDR boundaries according to Kabai and Cheihi* definitions.

Accordingly, in some embodiments an ami-f'5a antibody or end go re binding, fragment thereof described herein c;·.·: comprise a Kabar-defined, a ('hrChinoiebned. ora combined Rabat-Chodda-defined by hi chain CTh< set oboenod trom a light ehmn pelypuptide comprising 1 he amino acid o-queues ck-picied in any one ot bbgi ID Nigs; IV, 3~. or 43'. hi some embodiments, an an-i-Cba antibody or anbgen-buiding iragiueid then. ;)h described be re in can esenpriu: u Kahaudebne-j, a Chodtia-deluied, or a comb-ned KubauOmilm·,-defined heavy chain i'DR \ci obUim-.-J tr-.>rn a iv..-avs ehaln polypeptide comprising the amino acid sequence depicted in. bE() ID Nit 3“ -.a 45.

in mmi emhpd i metitg, tta aptbC'Sh anfibody described herein oompmes a bgM chain variable region connimnlrtg one se more of' a light ciirmi c'DR I c-.>mpri:fni.· or consisting of the foi lowing ammo acid set;uence: R.-\SES\'DS5'GNSI;MH (SEO ID NC'Ghbv a light >.n«in CDR 3 comprising or Csmrktmg of the ft>lUurmno acid sequence: R.ASAl l-S {RED i'D ND: 21). and u ligh· chain CD Rb oornpnsing or consisting of the following cunino acid sequence: vQRNRDIR'T (SEQ sD NsXodb !n ••ome embodiments, ar; anti-Cba antibody desenhed herein comprises a Hele dram variable region eossralnmg each of a iigbi chain CDR 1 composing or consisting of she lolkm ism amino acid sequence; R ASLSVDSYON'SFNHi (SEO ID Ν{.>:20:ο a light chain CDR2 comprising >>r e> insisting el rhe ioll< w me amino acid sequence RARN1 PS i RED ID \O'?. 1 I; and a Rein chain CDR3 ComprRmg or consisting of the f'llcn ing urn mo acid sequence OGSNEDRYT ·,.ΕΒΟ li.> NO;.).'.' e Rxs.mplary asm (Re antibodies comprising such a lich; chain vtinai.de domain include, e.g., the BN.I.o4, RNJ ’ό'\ β Y.i V/K BN,CE>6. BN.Dto-R and BNJehh anb-CRa antibodies described herein < CEt Ri//·,;: Table ID.

In so no embodiments, an anti-h 'G antibods described herein oooiprwes u iif!R elmih cafiablR region csasstahiibg: site ét more of: a light chain (RDM 1 eoisfrising or eon:· Dung of she Soihw. mg nntuv'add sequence" R ASCyVDS': GNSFiGR iSl-O ID NO DER a fight -.bain CDR2 comprising or convRnng of the IblRm me 3 mine acid sequence.. WAS! RES tSEQ ID NO. 3 K Land a licit· chain C. DR3 eon.ipyis.ing.or consisting of hie follow lug mnino ao:o r.,.-quenee: OO.RNEDFYT ·$Εγ) ID NO RED, in some embodiments, ;sn ;mtl-C?a antibody described herein comprises n Ugh; chain variable region comtdmng each of a EgSn chain RD.R 1 comprising of consisting of the lollop ing ammo acid sequence: RASESVDSYGNSFMD (SEQ ID NOE'OV a light chain C'DR a comprising or consisting oi the fbllowmg amino acid sequence: \YA STRES t BED ID NODS:, and a light chain CDR.'· comprising or consisting of the following amino acid .sequence. QQSNEDP't T CSEQ ID R:():.:.1 \. Fsempkirv anti-COa aniibqdisii comprising such aiighf chain viiiable domain: Include,: e<g,s (te BNJ3 71 and BND81 anti-Cba a nobed iv-- described her·.· it· π·ΛΛ· injnr. Table ID. hi some embodimenis. an am IR'5a anEbody described heroin comprises a heavy ehtdn variable region containing one or more of' a heavy chain CDR; comprising or Consisting of the following amino amd sequence' DYSMD ;RF.Q ID RO:/.b): a heavy chain CORE comprising or consisting of the follow mg amino acid sequence' \IN PNnGGTNY)K1FKD (SEQ ID Nf tD'-iy anq ;ï hcüvv chain (. DR s comprising or consisting of me following amino no id sequence: SGSYDGYYAVIPY tSff.Q ID RCb.'hn. hi some embodiment;·, an nnb-Coa antibmie deseribed herein comprises a heavy chain variable region containing es.cn of a heavy chain f DR t compiYnng ; -r e: 'n.hss :¾ of she foils >wing non no * ;d sequence. DYR VI D SFO ID NO:2HK a 2-,00. y chain CD if 2 c-nnpnsing os eoftsisiing of ’Ju.' follow nee ammo acid sequence AiMr.\^.iGTN\-NOK.FKD <SEQ !D NO.Df!: am! a heavy chain CDR3 comprisin'.’ oreons-sHne oï U;c loi lowing r;mine uoUi sequence. RC^VDCsYYAVlDY s$fQ i O K0‘30>. FNompUr. am: ('>;) nmihodies comprising sneb a heavy chain variable doneen ïnok;do, o.o., ;ho RNa/bd. BNj;q>7. BNJ371. and DNJY'h ;mn-f x; mthibodies

In some eed:xdiee.-eis. an asm-i’ '2.; unl-body described heron; comprises o heavy ohm π v mesbR ra o; o;; e<x2drung me or ns -re of; n heao y Chain CDR! composing or corssxhuu οί She following amino acid sequence: PYSMÜ 'Sir1,'; ID NOYhb a hex \ chain (, DR.: comprising <<r consisting of he; h-d· non nr am iso - as. id sequence. Λ1ΗLN 1 NY'S NN D vhahK.fi; RhR ID Ν0'4ό;; and a heary clnon CDRa comprising or corsahninc <-f the foilowhsg amino acid sequence: CdxRYRYRMDY (SbO ID NO:·;7) in some onihedimenR. as; ;mii-C.'5a ans-body described horem comprises a hear y chain van a hie region coniain;ma each of a heavy chain COR; composing »r smmosfmg oi She following ammo acid sequence. DYSMD REG ID NO.dbn a heavy chum CDR2 comprising an consisting of the Rakovam a; aha; acid sequence AiHtNTGYT NYNQKFKG iSioQ ID NO DC; and a heavy chain CD Re comprising air eonsixmg of the 'felovring annum acid sequence; GF\ DGYRRMDh tSEO iD NO.d'D. i:.\ernplar\ smi-Cbii aaubodics eosripnsmg each a heavy chain variable domain include, as;., Uu.· BN.D-hri BNjabG, BNJoHi. and BNJ3S3 amariha snfjbödiês <tesdrïb®di Jièfêlö; (WM mri® h Xlfclé .3},·

In some embodiments. un ami-€5» imbiamly or antigen-binding iragmerd mereol described heren; ran contain a be;;'·, a chain CDRc region comprising -fur ammo acid .sequence: APn PNSGGT NYSQK.RCDiSEQ ID NOYG. for example, as; antiiCêa liutibgcly described henrid can compile a hes&amp;><- ichai» -Yafiaite regi&amp;n: -eorUakung one or more of. a here, y chan; (' DR \ comprising an consisting of dm following atrtino acid .sequence·: DYSMD tSh-Q ID NO:28s: a heavy chain CDR3 comprising or c= -nCt-t-ng of the following ammo si·;, id sequence R1R ivN ROOT NY SO K FDD iSty; TD NOahTy ami a heavy chain ('DR3 comprising or consisting of da; following arrsisn; acid sequence: SO^YDG't'> AMDY iNF.O ID N0i3§); id some embodiments, an ^#tib«dy :d^«Srai?e4· tech* comprises a here, y chain a enable regien ea-neening each of a heav y chain CDR I comprising or eensbmng of she ieik.ovlng amino acni sequence: ON SMD (SiilO UN ND;2Rj: a heavy «ito in: €rDR2 temgttei 'hg; êt ébteteihg :óC tlie £é I Ictei hg amïte teid ste^^tesi A l N PNSOf ΠΓΝ YSQKFKD iSFQ ;D Uö:ó7;; and » hetn y chain CD Re comp-tehiio or consisting of the following amino acid sequence: SGSY1X5VY AMDY (SEQ !D NO 'U.SK Ac example of an anri-Cte antibody describe-..! herein, \\ibeh contains such a te&amp;w gtehi Itelypeptide and bhtte # fciffias 135a wtefi a Kd that ia Isas than 1. nM lx Use ten 10 IMS antibody described hde'-v.

In sonic embodiments. an anti-C te antibody er nmigonAhmiiny tragi-tont bvc-'e-it vie". pbee herein ee-malne ene el the exemplary light chain CDR set and heavy ho;n Ctey 'k· pa hinge i te -I dep-eied in I able i. .......

* " ‘ ;

The amino acid 'oqtienoos represented by the Si:O !'D MiX In Table ; are sot forth hi Tabic a

In some embodiments, she ams-Cte antibody decs not comprise exemplary CUR pairing e. lu «ome embodiments, the anil -C5a antibody is not BN.U~ I. in some embodiments. The light chain polypeptide of an anti-(.".ha antibody :described .herein can be-¾ A light chatttrpbiypspttde: te-g.. e fully: hitman xtCtemxnleecI % liglh dhaiir ppfypepTye}. $a yom®. epibodirsenty, the light: dMIn, polypeptide of’ atr ansl-Cte antibody described herein is a s light chain ixdypcpede -e.g., a Uiify human orhcmMiteif: te light team. pclypeghdeh The ammo aeld seriheneea of mimemuis light chain pel-peptides te.g., unmetous human light chain polypeptide') a?c ''xei.l-knpwn in the ajt &amp;ndect fortblii'i, erg., IC&amp;tetet ale tl:99:1),ysgjmi Exemplary telight chain polypeptide amino acid scqnenceN are am tbnh in Table a. in some embodiments. an ant· (’ 7 antibody described her-un can comprise a Hah: chain constant region, Fur example, fhe light a bain a·. annaat region can be a ,-light chain polypeptide com-tam reason o κ light ohuin eonsi-an man a;. The nmino seni sequence for a nun-be" •fhuman λ a an κ light chain eonsn-oU regie-no arc knoy, n In the art and described in, c.g., Kahm et ah i '·, sy/uv Fvemplaty ·/ light eh tun polYpcptide untino acid sequence:'· are set lord: In Table 2.

The heavy chain pub, peptide can comprise a. constant reel-.at (e.g.. a hete. y chain eonsSum region i tCH lt. heavy chain constant region 2 ;OI2/ heavy chain constant region a {'CUT;, uTtcitU chkm constant region 4 <CH41. of a eomhinutkni of eat- comprise an rc portion olasti nmnimuglobuim m<decuic. Tier Fe region can he, c.g , an he region from art IgGf igi.,1.:, by/a. !gG4, IgA. igM, IgL or IgD immunoglobulin molecule o? a combination of yon tost:·. ot'eaoli ol these. To be eienr. the a i tt I -1 ’ 5 a antibodies described herein can be, ,.-.g . ot igCli. igG.T igOa, igG-l. IgA, Ig/vl, iytf. or fgl) isoiype The arnica· arid ypc|tióoèès: for-# immbbr of buman heavy ehg® cohltani regions: are: IknoAtt fa the att Md descifb/d w„ i/g,,, irate et af. f I f 91),Αφο/.

In some ernbodirneuts. the Itemy chant pair peptide can cornpfiA a hybrid constant region, or a portion dte-eof. such as a (id? 04 hybrid constant region (see c.g.. Burton et aj. t 1/42'; .-/7 /comm /]. .Μ X; (no-babi et al. t, t °* > Ï t ,/£-,?* ./0-/ Γ-7. j-jsa- Μί©·^4·ίι·'®2)· Oor eyaigpieiand in :teeou.-;me<, ά sth Km hat numbering/ rise IgCli and igG4 constant region., comprise G Am .j. rcs-duoa rvi:c:aca the lyG2 eottsium region doos no? comprise residue 24/, but does comprise G,-<t. in. ,·. <72 04 itxbr-d const,mt region, \\ here the 2 -/7.70 region corner trom the (7,- r-cqnonce, the sonsfnm region can he further mod tiled /1 introduce a ubeme residue 7 ροχίΤη 2-io -o produce a t Ο ί <4 fum-u tneemg t ί :,-., /une; constant donum hybrids that comprise Cpw G.· ,.e;m tdso tie pan of engineered antibodies m a-xanc-incr v-ith the iKeiusnfè bsempjun hex, \ chain polypeptide ammo acid sequences -.ire set c-rrh m fable a.

Inc mut-C' aa am-body earn be. e.g,. one of the specific anbbodie*' e A-mphilcd in die xsotking examples. Bb/o/fT BNj.vS?. B/jSCg. BN.tfbP BNJTT-y RNjnKT 87.1371. or BNjpXi. The amine add sequences for these exemplified and-/7¾ an bio-die;·, v, htch can be used in cosy us tedoa o nh any of the methods described herein, sue set tort;·; In Table 2.

........................................................................................................................................................................................................................................................................ "Ab'5 ία Jjbk 2 icïc·:- u> rise alpistmimerpilerf^^ c-mmi'iks.

In sumo embodiment, mi ami-Cia antibody described herein comprises a C hoihia>dcftncd light chain CDR «er or a combined Knbut-Chofiiia<d£fincd light chairs ("DR set obtained from any of the light chain variable region* described in Tables 2 ot'%. In Nome embochmem, an mm-05 a antibody. or antigen-binding fragment ? hereof* described herein comprises a Choihia· defined heavy chain CDR set or a combined Rabaty Choihia-dctmed heavy chain CDR set obtained burn any of the heavy chain variable regions described in Tables 2 or 3,

In preferred embodiments* an anti-C5a antibody described herein binds lo CSa, but not to native, fuli-length C.\ in some, embodiments, an anii-O'u amibody binds 10 C5a, inn does not bind to the alpha chain of unc leaved* native 05. As used herein* 'uneleas od C5" refers to a OS protein that has not been cleaved into fragments C5&amp; and C5b by an AP or CP C5 conveiiase. An exemplary amino acid sequence for a human 05 ali'ha chain is set forth in Hambma os, ai. (1991), -mo/·.·;.. die sequence of which is incorporated herein by referenoe in its entirety* j.n some embodh.nentSv.au atni-C3u an tibody described herein does not bind, to pa? «dogs of human CS such as human C3a or human C4a.

The disclosure also features antibodies that erossbloek binding of-in an to 05 a amibody described herein t'e.g.. erossbloek.* any ooc of BNiJ3i<4. R\j36'·. BNJ37M BNJ300. BNJ3o9* BNJ.PL BNj.=5HI. or BMI.M3). As used herein, the term '"erossbloek ing antibody1' refers to an antibody that lowers the arnouri· of binding for prevents hi tiding ? of an anii-C5a antibody to an epbopeon a complement component C5a protein relative to tire amount ofbindinu of the ,mil-C5a antibody to the epitope to the. absence of the erossbloek iny antibody. Suitable method* for determining whether a fust antibody erossbloek.* binding of a second antibody to an epitope arc known in the an. f or example, croasblockmg antibodies eon be Identified by comparing the binding of the 808364 monoclonal ami-C$a antibody in the presence and absence of a test antibody Decreased binding of ihe BN.!3e4 antibody m the presence of the lest amibody us compared to binding of ihe 8Ν.156Ί antibody in the· absence of the test antibody indicates the test antibody is a erossbteekiog airiiteby., in some embodiments, die binding of no antibody m C5a can inhibit the biologies! activity of C5a. Methods for measuring 05a activity include, e.g . chernotaxis assay-'.. rsdioirnmunoa<%ay s (RIAsT ar enzymc-linked in)muno.spccific assays tELIS.A) (.vee, c.g.. Wam and Zvuiflcr t k)7 1) J CVb; /m.yv; 5Q{3 ):606- 16 and Wutvner et al. t1 00i { (y.-fiiptL'iin-rJ injliiftnn bwOWIMb). |n scene embodiments. tbc binding of an antibody or antigen-binding iragmetO thereof to f'5a can inhiba C.'ha-rnedimed ncuOOphd aetbabcsn in r/m<e Sc liable meebode tor determining o Her hor un aml-Oe am i body inhibits Cé», mediated neutrophil aedvaston ia exm, or the extent na vrind; tbc antibody mb baits aciisation, ara known in the an and exempldled In the working examples below. For example, human neutrophils obtained from healthy donors cart be isolated and contacted w I ill isolated human tffa in the presence or absence of a tees anh-Cëa antibody. C fa-dene udeut activation of human neutrophils can lx- measured as a function of mydofarcsxlebsc (MPOi release from the cells In the presence of C5a. An inhibition of the amount cd MPO rdeased front the cells 1st the presence of C5a and tin.· test antibody, as competed to the amount of MPO released from cells m the presence of ( 5a and a control antibody, indicates that rhe test antibody inhibits C da-mediated neat tophi! activation.

In some embodiments, an anti-i 5a antibody or antigen-binding fragment thereth does not inhibit gw docs no? subsume ally inhibit) the actobiy of complement component fbb as core pared to the Ic^ci of inhibition {if any) observed by a corresponding control antibody or :.m;iwwd>mdmg fragment thereof it.c.. an me-body that does not bmd to tree (;5a or Cbj, f'5 actix Ity can be measured as a function of Us cell-lysing ability m a subject's body fluids 1 he cell-lysine ability, or a reduction thereof, of C5 cart be measured by method·.; well known in the art such as, bar example, by a conventional hemolytic assay such as the hemolysis assay described by If a leas and Mayer peris). “Experimental I mntunoc item istry, 2“': Ifdfm.mf' 155-240. Sprint:lick!., IL. CC Thomas (1061;, pages 155-130, rtr a conventional variation of that assay wteh as the chicken erythrocyte hemolysis method as described In, e.g„ Hi Urnen et a!.. (20(61 ί .V Engf J a A A 351)(61:552. bt some embodiments, 05 no tie ity, or inhibition thereof. Is quantified using a ('fiéOeq assay. The CliéOeq assay -s a method for measuring the total classical complement activity in serum. "This test is a lytic assay, which uses a-ttibody-sensitixed erythrocytes as the activator of the classical complement pathway and venous dilutions of the km serum to determine the amount required to give 50?'« lyses ((/1150),. The percent hemolysis can ho determined, for example. using a .spectrophotometer The i'RbOcq assay pro\ ides an iocliccct measure of terminal corr^tlemoru complex ΠΌ?;· formation, since the '1 CC themselves are directly responsible lor the hemolysis that is measured. fhe assay is well know n and eommonly practiced by those of skill in the an, BrieHy, to activate the elas-ucal complement pathway, undiluted sewun samples (e.g., human serum sant pies) are added to roicroassay weds eomammg the ant t iaody-sensitised erythrocytes to thereby generate TC'C. Next, the acnsvued sera an; diluted in microaxsa} v, eiK, which am coated with a capture reagent (eye. an antibody that binds to one or more components of the TC'C) flu; TC'C present in the activated samples hind to the monoclonal antibodies coating the· surface of the mieroassay wells. The wells ate washed and, to each well. Is added a detection reagent that is detectable bee-led and recognizes la; bound "ff C The detectable label sou be, eye, a fluorescent i«b>„ I or an enzymatic label, ihe assay results are expressed in f H50 unit equh aletns pet milliliter (CB50 U ho ntl!.

Addiüdn^''m.Mhö.d-S':f®r defecting tnd/or measuring €S activity m viim are set forth and exemplified in the -corking examples

In some embodiments, the buiding of nu antibody to C5a can inhibit the interaction between C'5a and C5aR 1. Suitable methods lor detecting anchor measuring the interaction henveen C5s and C5aRJ tin the presence and absence of an antibody? arc know n in the an and described in, e.g., Mary and Boulay i 1995) EueJ Haematol 51(5),282-287: K.anekoet ai. (1995) *6« 1 >:I49* 154: Giannim ct a). (1995)./

Biol Chetn 270(32):19166-1^3 72; and U.S. Patera Application Publication No.

20000160726. For example, the binding of detectable labeled (e.g., radioac lively labeled) C5a to (75aR t-vs.pressing peripheral blood mononuclear cells can be evaluated in the presence and absence of an antibody. A decrease in the amount of delectably-labeled €5a that binds to C5aRI in the presence of the antibody, as compared to the amount of binding lu the absence of the antibody.. Is an. indication that the antibody inhibits the interaction between C5a and CSaR I hs some embodiments, the binding of an antibody to C 5a can inhibit the iistcriiciiou between C5a and ( .51.2. Methods for detect me and or measuring the interaction between (35 a and (551..2 arc known in the art and described in. c.g . Ward (2009)./ \f-.tl :bW H?i4i:375-.v>: and Chen et at ¢2007» \’anjn> 446(7132):203-207. Addition;·.! methods tor evaluating the biological effect o; an anti-OSu antibody described herein are exemplified in the working examples below

In some embodiments, the mnl-Cba antibody specifically binds to a human cornpie.ment component (35 a pimcin (c.g., the human (75a protein havtng the amino acid .sequence depicted in SEQ ID NO:; or SEQ ID NO:2>, The terms "specific binding."' Specifically hinds." and like grammatical terms, as used herein, refer to two molecules forming a complex (c.g., a complex between an antibody and a complement component C5a protein) that Is relatively stable tinder physiologic conditions. Typically, binding is considered speetdc 'Alten the association constant (kn is higher than 10'1 M ss *. Thus, an antibody can specifically bind to a C5a protein with a k:, c*f at least tor greater than) I01' (c.g.< at (cast or greater than 10’. 10'\ 10'. I0"k !0:k iOW 10’ . 10 k or 10’''or higher) M ’ksf. In some embodiments* an anti-Cba antibody described herein has a dissociation constant (ktl) of less than or equal to 10 (e.g., 8 x 10 ", 5 x 10'\ 2 x H5 \ 10'1.. or 10) s'" In some embodiments, an ami-C5a antibody described herein has a K-> of less than !0'\ lO'k KW'3 i0:\ or 10 " M. The equilibrium constant kn is the ratio of the kinetic rate con sum Is - W I·..-,. tn some embodiments. an ;uni-C5a antibody described herein has a K.;, of less than 1.75 x 10'' M. Examples of ant t -C5a antibodies rltat bind to C5a with a. K:; that is less bum 1.2.5 x 10s M mcSude. e.g.., the BNJ3M. BN.I367, BN.137L BN.I3/H, BNj366. BNJ.W9. BNJ3H1. and BN 1383 ami-Cda antibodies. irt soirtc embodiments, art ami-05a antibody described herein has ;s Ki; oflcss than 1 x 10'1 M. Examples of anti-Cêu a mi bodies Hon hmd to C5n with a k;> that is less than 10" M include, c g.. the BNJ3M, BN.1367. BN1.V78, BN Bob. BN.i3o°, BKDHI. and BN.I383 anlt-CSt; anttbodies. in some, embodiments, utt ami-Cia antibody described herein has a Ki; ol'iess than 5 x 10 1' M. Examples of ami-C.5a antibodies that hind to 05 a with a Km that is kss than 5 λ 10’is! M inehtde, e.g., the BNBb7, BNB?k BN ?3(W BRBiW, BNBR. and BNJ383 aori-CSa anttbodtes in somt; embodiment, au ami-C5a sou body described herein heus a R;: ofK'Sa them 2 x 10 t-) M. Examples of aiui-C'da antibodies ids; [end 10 C'5a with a Re that is less than 3 \ ;0 !'! M meb.ide, c.g., One BNjah'y BN j 3 ót\ BN i .9-9.. BN.mbR oud BNiJnHd ami-E 5a amua'scnex.

In some embodiments, ait ami-Clkt antibody described herein lias a Roofless than I x 10 M. Exusnplos of ami-OSa and[00:.0os Oku hind io 05a with a K.e that is teas than 1 x i0'i:: M include, e.g,, the BNBiY), BNJdSi. and BNJ383 amt-C5a amihodies.

In some embodiments. ;m anti-05« loibbody described herein has a Ivooftess than 7.5 \ ICO' M. Examples of anti-C da antibodies that bord to CBa with a Re that is less than 7,5 x HO' M include., e.g... the BNjadd and BNjRan ,ιηιΕ05η antibodies

Methods Γογ determining tit affinity of mi ami body fora protein antigen are known in die art. Tor example, die attinhy of an antibody tor a protein antigen can be quantified name, a sauieit of techniques such as but not limited to. Western blot, dot bloi; Biolayer hsteillrometry. Surface Putsmon Resonance : SPR! method to.g.. Bl Acosv system; Pluinriaeia Biosensor AIR Uppsala. Sweden and Piscatasvay. N 3 y or enzyme-linked im?minosneeiln: assays HiiiSA). .See. e,g., Harlow and Lane {P>bM “.Anybodies: A Laboratory Manna!5' Cold .Spring Harbor 1. aboratory Press, (.'old Spring Her hot, N.Y.; Benny K... Lo {200-1) "Antibody Engineering. Methods and Protocols." Humana Press AS BN' I 58P29092 \ χ Borrebaek {1002) "Antibody Engineering. A Practical fittide." \Y.R. Freeman and ( Ό., NY; Borrebaek {100*5) 'Antibody Engineering," 2lui Edition. Oxford l adversity Press, NY. (Oxford; Johne cl a!. {;093)./htwumt··! .tƒ*.·//; .1.09 PAN W; jo assen et al. 1RNU) .-ive Biol CC n Μ: ί°-26; and lonsxon et id. 11901} dm<V;7mR/m'x It:b20"627.

Any of the light' chain COR sets or tight chum variable regions described herein can be paired with any of the hcuv> chain (OR sets or heavy chain variable regions described herein, h is well within the purview of die ordinarily skilled artisan to, c.g.. confirm {test) that an anti-BSe antibody generated bs such a pairing possesses the desired affinity or activity Suitable methods for confirming the activity and or affinity of art anti-f'da antibody are described herein.

In some embodiments, the amUC'da antibodies described herein bind to both human C'5a hCbbys and C5a trout a rton-human mammal suds as a nort-hunturt primate te.g., cynomoigus macaque5, In .some embodiments, m anh-OSa antibody o> anttgen-binding fragment thereof described herein does not bind tu para logs of human C5u such ,ts C 3a or Oh born the same non-hunnm mamntaitan species.

In some cnnxMiments, en anti-CSn antibody or antigen-binding fragment thereof described hereni binds to bee hC5a and a cynomoigus macaque i‘5a protein αηηρπ-brtg. or constating oh the lolloping amino acid sequence' MI.QEKIïbl\ AR VK.HLVVRK. CC WXi\ RING OFVCfcQRAAR ISVGPRCVKA FTBCCVYASQl R ANN.SHK.DtQi.GR tSBQ ID Ni.)-17PI In some embodiment*, an aoti-Cêa antibody or antigen-binding fragment, thereof described herein binds to tree hC5a and a rhesus macaque 05a protent comprising, or consisting of. the amino acid sequence depicted tn Sr.Q ID NO: 17-). bn some embodiments, an antibody, or an antigen-binding Fragment thereof, can bind to a desargmatod form of C5a protein from a non-human mammalian species t o g., a nom-human primate species's, for example, the antibody or antigen-binding fragment hereof can bind to a bee L.5a~desarg protein from cynomoigus macaque or rhesus macaque, the protein comprising, or consisting of, the following ammo aetd sequence: MLQfK.IEEIAAfCYKHLVVK.K CCYDGVRIMH DBÏGBQRAAR ISYGPRC'YKA PTPCO V S' ASQi .R AN NS I IK DLQI .0 ISF.Q ID NO'IMib

In some etohodlmems, the ami-CN) antibodies described herein bind to mouse C!5a li.e., the tree C5a from mouse}. In some embodiments, the anti-Cya antibodies described herein bind to mouse f'3a, but not to human C5a. In some embodiments. an aoli-CSa antibody described herent docs not bind to undcaved, native {fully-folded) m(>usc C5, In some embodiments, an and-C5a antibody described herein does not bind to para logs of mouse C5a sucb as mouse €3a or mouse C4a.

An anti-mouse CSa antibody, or an antigen-binding fragment thereof, can hind fo a mouse €5a protein comprising, or consisting of, the following amino acid sequence: LiGdl<.il:iiTMAKYKilSVPKkCCYIXY\RVNFYFKY-BR VARYTIGPl ORAFNECCT IANKIRKE$PHKPVQI..GR tSf.Q ID NO'S i}. See also, c.g , Vvetse! ct al. Π9Υ7} BitKhew 26,737-743. In some embodiments, an ansi-mouse CSa antibody, or an antigenbinding fragment thereof, can bind to a desarginated form of mouse C5a protein eon-prising, or consisting of, me iblkvo mg arm no acid sequence: ERQICfEEQA AK V KJ IS VPKK.CC Y DG ARV N ΓΥ ETCOIRV ARVÏ IGPLC 1 R.A PIS ECCT lANRjRKESPHkPVQLG sSEQ ID NQ:S2). In some embodiments, ihc a mi-mouse CS&amp; antibody binds to bosh the full-length mouse C5a protein and she dei*ijtuted form of she mo use (. 15a protein.

Aaanti-moo.so C 5a antibody described heroin easy e.g , contain a hghr chain CDR act obtained front a light chans variable region polypeptide comprising the following amino acid sequence- nYLTï^'sPAfMSASLÜEiKYTMSCRAS.'SSVNVIVWYQOKSDASPKÏ.WiYYTSNLAAF G V ΡΛ RFSGSGbGNSYM. 11SSM KG PDA AT Y' YCQQF' PSS PLTFG VG' ΓΚΧΥ l K.K (NEC? ID NO:53f. For example, ast ami-mouse C5a antibody east contains si) a RGba'-defmed sight chain CDR I com pricing, or consisting. of. she to! loo nsg amirut aesn sequence· RASSSYNY1Y (SEQ ID NO'54}' (il; a KaKn-deflned light ehahs s'DR l comprising, or constNong of, the following amino acid seqsienee' YTSN1.AF {SEQ ID NO;55K and'os siii} a Kabat-detmcd Hunt chain (OR) comprising. orconsisting of she following amino and sequence. QQFTSSPLT {SEQ ID NO.êbt.

The ami-mouse C5u antibody cast contain a light chain coststasts region, e.g , the mousse kappa light chain consiant region comprising. or eosssistsfsg of the follows sty ammo acid sequence: A D A APT V S i F PPS51 EQLTSCiG A S V V C'F LN N F Y PK DIN V' K W KI DGS E R QN G Y LNS \V TDQDSKJDSTYSMSSTLTLTKDEYERHNSYTCR.-M'HKTSTSPI VKSFNRNEC (SEQ ID NO :5 7). 1st .some embodiments, an ami-mouae C5a antibody described herein can contain an amino-terminal signal peptide, e.gm a signal pepneie comprising. or consisting oil the ibUowmg ammo acid sequence. Y1GWSC! I LELY ATATG VH S (SEQ ID N0.5M. in some embodiments, an ami-mouse C5a .-Antibody described herc-ist cast cons^in a bght chairs polypeptide comprising, u; eor^istirsg of, the follow sstg urssisto acid sequcstee' P EI VL"i QSP AI MS AS 1 .GE K V 7 MSCR ASSSY NY 1YWY QQRSDAKPK LWÏY YTSNLA PG VP A R Γ SGSGSGN S YSLTISS M P.GE DA AT V Y COQ FTSSPIT FG Y GTK LIIEK R A D \ \i'l\ Mi PPNM Qï lMw< W\ \U1 NTs b HJYIW KA\ KIIHsSs RO\tA 1 NSW ,|i QDSKDSTYSM SSTLTI.' Γ K.DEY E REINS Y IC RATH KTS' PSP IYK SFNRN EC s SEQ ID NO: 5k) or MGWSC! i L i< l V .YÏATG VH SR &amp; ï V L I'QSP A IM S AS Ui FK.VTMSC RASSSVN Y IY W'Y QQKSDASPK L W iY YTSNI .Λ PG V PA RFSG SGSG NS N 'Si .Ti SS M EGPD A A ΓΥ Yt.W TSSPLTPGYGI K L 1:.1.. K R AD AAP iYSIFPPSSEQL rSGGA S Y VC FL NN ΓΥ PK .DIN VK W KI DGSf: RQNG V ί .N S WTDQDSK DS T Y S M SST l 11IX PF Y f: P Η N S ΥΤί,Έ ATKKTS TSP ί V K $F N RN FC (S.F.Q Η) NOYO). In some embodiments. an anti-mouse C5i antibody described herein contains a Isgbs chain poiypcpdde comprising ammo acids 2 so 2 H of SU) ID NOYY In some embodiments, an anti-mouse CSa antibody described heroes oosttatna a sight chair; polypeptide comprising asnirio seals I ίο ί P ami 21 ιο AG of ShO ID NOYi).

An anti-mouse t 3a antibod}·' described herein can, c.g , contain a heavy chain ( DR set obtained from a heavy mmm variable region polypeptide comprising tire to doming amino acid sequence' f.i'YQl.GQSGPn.YKPGASYKiSCKASGYTFTnYYYiNWYKQSHGKSLFWIGYn'P NDGDTNYNOKFKGKATLTVDKNSS'IAYMELRSLTSEDSAVYYCAR pyysdyom DY WGQG FS YTVSS {SEQ ID NO.61). for caarnpkv au mni-mccmsc C cm ton;body can con sain: (it a Rabat-defined heavy chain CUR I comp do my or consisting of. the following amino acid mqueocc; DYVYIN (SU) ID NO.oa); {sit .a Rabat-defined heavy chain CT)RC comprising, or consisting of, the toiler isig ammo acid sequence. \ iYPNDGDTNN NORFlvG (SU) ID NO.bc): anchor f-hi > a Kabas-dcfincd hemey chain (1DR3 comprising. or consisting oh the following amino acid sequence: PYYSDYGMDY ISEQ ID NO Yd}.

The anti-mouse C5a aottbody can contain a heavy chain constant region, in some embodiments, an unri-mouse (,'5a antibody described herein can contain an amino-terminal sigsval peptide, cm., a signal peptide comprising, or consisting oh the ibliovvirig amino acid sequence: MGWSC.'llLFLYA'T'ATGYHS (SF.Q ID NO.oSï.

In some embodiments. an ami-mouse C5a antibody described herein ear; eontain a heavy chain polypeptide comprising, or consisting of. she following amino acid .sequence. L EVOI..QQSGPFLVKPGAIsVK!SCKASOYTFTDYYYlNWYKQSHGKSLr.WlGYIYP MYdGYh V)M Mik Ml U DKn<G I YMÜU1 ) \ | DN \ A } ( MGGbSD'UA; DYWGQGTSVTVSS (SF.Q ID NO;tY)or

ViGWSCtILFI..VATAI’GVMS 1..FvQLQQSGPELYKFGASVK1SCRASGY ITTDYYYi NWVKQSHGKSIJiWiGyïYPNDGDTNVNQKFKiSKATLTVDKSSSTAVMELRSLT .SrDS:\VY YCARÏ^YSDSTjMDYWGQijrSVTVSS iSIrQ ID N0:f«7i \n some embodiments, an and-mouso v 5a antibody doodbed herein contains a heavy chain polypeptide comprising amine acids 2 to Id 1 oi'SIiQ ID NOh-o. in some embodiments, an ami-mouse 05&amp;antibody described herein conrairuvu homy chain polypeptide camprDiug urmno semis I to id and 2! io 140 or'ShO ID NO;n7 in some embodiments, ait mynmousc (.'da antibody described herein contains a heavy chain constant region polypeptide comprising one or more amine» acid substitution'* from the above described sequence.

In sonic, embodiments, an arm-mouse ('da antibody described herein contants .» tight chain polypeptide composing. ti) a light chain COR] composing. or consisting ob the annuo acid sequence depicted it» SFQ ID 'y(vy4; <n s a light <.hain CDR2 comprising, or consisting ob the annuo aetd sequence depicted ut Sf O ID NO'55; and lm) a light ehatfi ('DK3 composing, or consisting ob the annuo acid sequence depicted in ShO ID Ni f 5u< ι y} a hero y dnun OPR t composing, or consisting ob she amino acid sequence depicted m Si::Q ID bO.'tbb u ) a heavy chain (' DI\2 comprising, or consisting ob the amino acid -Cijiteuve depicted hr SI:Q ID NU.mb and <» tvit a hea\\ chain i.'DRo comprising, ett consisting e»t, the arm no acsd \equem.e depleted is» SbQ ID \0:t>d

In some embodiments, an amwnonse 05>-, anti bods described hcrom contants a ttghi chain polypeptide comprising, or consisting of. the amino mid sequence depicted m SFQ ID \('y and u heav y chain polypeptide comprising or consisting of. the ammo acid sequence depicted in SEQ ID NO:0b.

In some enstx«diments, an amt-05¾ antibody ^gserti>ed;;ksreiji-.-eitd bind to bemad (25a and to mouse CSm λ sJ. ΙΛ .-.t d.ïS.. 0 2. b..! .CD.s Id /d .Λ /.1 (2. Γ.1λ>.·. .. ut.t dij .r (:. !.. Λ Λ h"; v>oi.t c s.. ry.t e!.. A ry r«g:y.. y jD y tyi i. t u\. .1;. y; t ΙΛΓ:Λ0Λ>.1 .et. .1. .1 l.O.V.S'dè Λ

Th c disclosure also feature-, methods lor producing any of the autei Ό anth»odles or antigen-binding fragments theteoi described herent, in some embodiments, 'methods tor preparing an anybody described herein ecu include immunizing a subject -mg., a nonhuman mammal) with uu appropriate immunogen. Suitable Immunogens for generating any of the antibodies described herein are set tenth herein. For example, to generate an antibody ihai bind1* 10 05a, a ski ik'd artisan van immunize a suitable subject fo g., a nonhuman mamma! such js a rats a mouse, a ger'bil, u hamster* a doe, a oak a pig. a goat, a horse, or a non-humtm prunaie) with a full-length C da polypeptide such as a lull-length 05 a pols peptide comprising the amino acid sequence depicted in NkQ ID NO, I or the desargmated torna of C5a (e.g., the human C5a desarg comprising. -he a rot no acid sequence depicted m SFQ ID NO;.:), hi sonic embodiments, the non-human mammal is ( 5 deilciem, e.g.. a f.'5-Jeficicm mouse, described in, e.g,. Levy and kadda (I'TO } \or w n- Bio; 2:00.).5 Odd; (.'mdoar et ai. i ITM)./ Om (NrDd 21(2). 122024: \\ else! el. al i i 990),-^ Βίοι Chens 2o5;2455-2-1 a0, end Jungi and IVpys ι 198 i) /nmmmk >01 95;2):271 2'? 9, Human C5a can be purified trom human serum as described ou e.g.. \ki "arth> and ! lenson i 1° k>),/ hniuunoii 123<b) 251 0251 ? and Manden no ei ai. 1 0.02),/ /;?ί>ί>ηη·>/ \ii r/a>,·Α .55.0.):41 -50 Nee also the working examples. Human C 0¾ can ulso be generaled. O ruVv; as desert bed in, e.g , Vailota and Midler-kberhard {I -0)}./ /ο·;.· ,t/cJ jjj. 1109 Ponded human (25a is also commercially ova.dunk trom, e g.. Complement Technology. Inc. (catalog number Λ 144; Tylen Texas) Recombinant C5a can also be generated by one of ordinary skill in the art as described in, c.g„ I uTc el ai,· 199···!} /0-..y; -/ ;t-t tap. 1108. suitable subject te.g., a non-huniun mammal) can be immunized with the appropriate antigen along oho subsequent booster immunizations a number of limes sufficient to elicit the production of an antibody by the a/amman T bc mtrmmogen can be administered to a subject {.e.g., a non-human mammali with tut adjuvant Adjuvants useful in producing an antibody in a subject include, but arc not htnited to, protein adjutants; baeterud ndluvavus, e.g., whole bacteria iBCG. konne/mr/cr/iim nunn/m or Sahn<iiti Ho miofu:-,<*/;) and bacterial components including cell wall skeleton, trehalose dimyeoiaie, monophospbory! lipid Λ, methanol exitaemblc residu·:· (MFRi of tubercle An7//no, complete or incomplete Freund V adjuvant; viral adjuvants; chemical adjuvants, e.g., aluminum hydroxide, and uxloscetute and cholesterol hem (succinate Other aditixants that can be used in ihe methods for Inducing an immune respost.se include, e.g. cholera a win and parapoxt tins proteins. Nee also Biegci ad, ι1999) Auroimwamfy 3 ; 15-2-1 Nee also, e.g., i.odrueli ei a I (:900- l-'m a 00 j.k: 1059-; Oho; Johnson ct a! (IWiJ Mol Chvm 42:4640-4^49: Baldridge et al. (llWt Mninnk: .10:10.^-10/, and Gupsa cs al. < 19°5) hierinr 1 3- Mr 1 Jóu-1 2';s\

In .some omhodintents She methods include preparing a hybridoma colt kite that ace ras ca a monoclonal antibody that hinds to the isnmunogest For e sample,. a .suitable mammal such as u laboratory mouse is IsumunMed with a Cl :3 a polypeptide as described abose Asnsbodwproduemg celts (e.g., B cells of she spleen) of the I nun uni.red masnmut can be isolated mo so sour days after at leunt one booster hruonniaaiion of the unsnustogen and then gnaw rt briefly in culture before fusion with cells of-t suitable myeloma cob line. 1 he cells cats be fused iss the picsenee of a fusion prontoter such as, e.g., vaccinia virus or polyethylene glycol. The hybrid cells obtained in the fuss on are cloned, and cell domes see re imp the desired antibodies arc selected. For example, spleen: cells of Baiba: mice ins run nosed with a suitable immunogen can be fused with cells of the myeloma cell line P \! or the myeloma eeH line Spa 0-Ag 14. After the fusion, the cells are expanded su suitable culture medium, which is supplemented with a selection medium, for example HAT medium, at regular intervals in ordes' to prevent normal myeloma cells from overgrowing the desis'cd hybridoma cells t he obtained hybrid cells are then screened for secretion of the desired antibodies, e.g , an antibody that binds to Ο5ο and inhibits the interaction between C5a and a Odu receptor ie g., 05uR Π.

in some embodiments, a skilled artisan can Identity an a.nh-(" 5a antibody front a non-imntune biased library as doses')bed ist. e g., 0 8, patent no. 6,?00,0on (to Knappik et aft Morpbosys AGs and Scimonbroodf ot at, (2:005s .VuMrk .iritis R<:> 3 A; A mb I

In some emborhmeats, the methods described herein can involve, or be used in conjunction with, e.g., phage display technologies., bacterial display, yeast surface di.spl.ay, eukaryotic strul display, mammalian cell display, and cell-hoc (e.g., ribosomui display) antibody screening feehstioues (see, e.g.. Mo et al. (TOO; ij Beamin' 183.092-1-Ob35.. Cornclts (2000) Gar Opiz Mmvr/wM 1..1.:450-454: Kiemm et al. (2000) V/n.-'V tbit >;<· >yy o4p :.)02 5-305 2: Kick:· et a I. {I (!0M PnAchi tiny j0; i .)0)-.1310: Yeung ei ;d :2002} Bimcchtioi Proy .1.8:212-220.: Roder et al s 2:000) \h:thuds Ertzyurdoyv 328:4.)(i-444; (jrubherr et at. (20(5 1) Owl· < 7?„ν« Hiyh Tiintxyhpn JSM'ee?·? 4:185-102: Michael et. al. (1995) Geur Tiur 2.660-008; Perr-boc-v et. at. (2001)./ Vind 75:7107-71 :.):

SchaOlrzol et ai (S 999s J ίααααααΐ SUïIvoS 27jg I 19-B5, and Hastes ct«I. (2000) Sa; Siotcdwol 18:1287- 1 292 s.

Methods tbr identifying anybodies using various phage display method- are Known in the art. ia phage display 'method1-. functional antibody domains arc displayed o» the surface of phage particles winch carry the polynucleotide sequences encoding them. Such phage can be an heed to display antigen-binding; domains of an a bodies, such as Fab, Fv, or disulfide-bond stahilked F\ antibody fragments, expressed from a repertoire or combinatorial antibody library (e.g., human or murine). Hinge used m these methods arc typically Filamentous phage such as id and M13. The antigen binding domains are expressed as a recombinamly fused protein to any of the phage cost proteins-ptlL pV'tif or plX. See, e.g., Shi cl at. -:2010),/,)/-9 .V,-*?.385--7uó. Examples ofphage display methods that can be used to make the hrunmtogiobtslhis. or (rag me ms thereof, described herein include those disclosed it! Brink matt ct al, : I 9°5 >./ /memm </ .) 7mm a A 1^2:41--50: Arnes ct al. Π 995}./ hvniunal Sk'thad', Lyse! 86; KmtdcbonHtgh et al. < 1994:- Ear J 1t /me >?;· > / 2 4; 9 ή 2-9 5 8; Pu··, sc m al (199?: G'mm j.8g;9~1 K; Burton ei al. i i’-b/bighema.o />? /mmmmégm mg 1i} -280: and Ph i publication nos, WO Of) 02809, WO 91 10777, W(> 92 01047. WO °2186|9, WO 97- U2M\ WO 95 15982. and WO 95 70401 Suitable methods are also described in. c.gc IhS. patent no.v 5,598,42(:: 5.77.7.409; 5.,403..-184; 5.580.717: 5,/7 7,90b; 5 .,750,?53; 5.0.3 i .0-17; 5,571,698; 5 427 908; 5.510,077; 5080,225; 5,o58m27; 5077.747 and 5,969.108,

In some embodiments, the phage display antibody binaries can be generated using mRNA collected bom B cells from the Immunised mammals, For mmmpie. a splenic cell sample comprising B cells can he isolated Iron- mice -.mm sun reel vein: 05a polypeptide as dcsciibed above mRNA can be isolated from I he cells and converted to eONA using standard mideeular biology· techniques. Sec. c.g , Sumb<ook ct al. 11989: ’'Molecular Cloning: A Laboratory Manual, 2"'' i:\1itiot:C' Cold Spring* Harbor Laboratory Press, Cold Sprutg I iarbor, N V . Harlow and Lane (19H8 ·, .m/mu Benny K.. C 1 o (200-1), ΜψΓι,\ and Bmrebauk {1995), .atpru. The eDC 4 coding for the variable regions of the hoary chain tmcl light chain polypeptides of immmtoglobtd Ins arc used to construct the phage display library Methods for generating snelt a library are described in. e g .

Mere. c; ai * ,/ Λ\;ί,'ν>·,,.·< Xfnt'oM peuk; 1 ! 5-T I>j Vsro et a; Code 5 l>V< ó-vn,/ èsStP? U.HK9 b^M and Fnghetg ai i \v95) λ,UW ibW ? ;. .)55-.U'o.

In some embodiments a i.orobirouion of sdeehoo and sereuhnc eau lx employed, ίο akaHifs au untihods of interest hem, e.g , a population of lobodomn-clèrked antibodieson<> phage dkphn antibody iiouey Snitabk methods arc known in da. an and ;nv described in, c a , 1 ioogenhoooi < oh*"') :r<.u«ó w ΛΥ -o... hn->h-er 15 o2-"O. Brinkman Cf ai i kJ;>ók 00--00 \mes et al < I-Uk scoor KeidehO'OUüh Cf al. 5 nemv. Pcrsiv ei al < i 99? a ---0-00:: and Burton et ak {1 ^1), o-gWi kor example, -s pU-raljfy of piuearrnd seaors, caeh encoding a rusion prounn of -.¾ baeienophaa:. coat pattern ie,μ., pill. pVill or pl.N of VI15 rha0,:) .md a different antUmi-cosobming region are nr-n.ieeed using standusd molecular too logs no luskiues and dan introduced snio a population of bacteria (o.g , /: r< -//) expression of tiie bacferioph.u’e sn bacteria ran. in some embodiments require m-c of a helper phage, in souse eoshminnems, no is.: I per pda nr :s required {sec. e g... Chasteen et ok C00o> A„v.\/, * >M -Vo. 59Ci).el95). Phage ptodixed froni ihe bm let 5a ,ih avows cd and thus eomaeled to, <. .0,, a liege* .onoo-n bound to a .sohd supper? *murtehikvdi, Phage ma> oko Pc cmUuded ;o antigen ni suhnion, and the complex is subsequently bound '0 n who support 1 i- sCuoo cufoorlunents. the innnofnuaed phage are five phage of interest. Aeeoi'dingiv the unbound phage ate removed hy oashinu the support holme, im: the naslt step, bound phage rue then cltued front the ---0:id support e u., using a Urn pj I buffer or a tree target antigen competitor, arid recovered by mhacring bacteria. In some embodiments, the phage that are not immobilized are the phage of interest. In such embodiments, the population of phage cun be contacted to the antigen two or more dines to deplete from the population any of the phage that btnd to the support Unbound phage are then collected nod used for subsequent -screening steps.

To enrich the phage population for phage particles that contain antibodies having a higher affinity for the target antigen (while reducing the proportion of phage that may hind to the antigen non-spec;deally), ihe eluted phage (described above) can be used to re-1 π feel a population of bacteria! host coils. The expressed phage are then isolated from the bacteria and again contacted to a target antigen The concentration of antigen, pH. temperature and inclusion of detergents Arid -adjuvants during contact can be modulated to enrich Coe higher arrmby ami body fbigments. The unbound phage arc returned by washing the solid support. The number or cycles, dura uon, ρί-K temperature and inclusion of detergents and adjuvants during washing can also be modulated to enrich: for higher affinity antibody fragments. Following the wash step, bound phage arc then eluted from the solid support. Anywhere from one to six iterative cycles of panning may be tried to enrich tor phage containing antibodies having higher affinity for rite target antigen. In some embodiments, a deselection step cun also be performed in toniinwiion w ivh arts oi the panning approaches described herein

Individual phage of the population can K isolated by mica mg huaerb and then plating at a density to allow formation oi'monoelouai antibodies.

For example, to identify using phage display techniques an antibody that binds to C5a. but not to F:5. the following panning approach can be employed The population can first be contacted to u surface containing hound name, foil-length human 05 The process can be repeated two or mote times, each time collecting the unbound phage. The. population can also be contacted to a solid support containing surface-bound C4 and or O proteins. Unbound phage from the foregoing steps are then contacted to a surface comaiumg bound C5o or de.vnrg mured Ufa Phage that land to 05a are eluted from the surface and recovered by infecting Uaeterta. Iterative rounds of phage selection may be performed. After one lo six rounds of seleumn, individual recovered phagemld can be screened for expression of antibody fragments with the desired specificity and affinity. A subpope UFon of antibodies screened us mg the above methods cun be characterised for then· specif cits and bieding affinity for a particular immunogen te cm 05:0 using any mmamoiogwai or biochemical based method Known m the ml. For example, specific binding of an antibody to Can, as compared ιο native. ihtMoi'gih C :5, may be determined for csample using immunological or biochenoxd bused methods such as. but not lirmicd to. an El. ISA assay, SPR. assays, imniunoprecspitaiiou assay, affinity chromatography, and equilibrium dialyses as described above, iromunoassays which can lx* used to analyze immunosncdl'k binding and cross-reactivity of the antibodies include,: but are nut limited to. competitive and noo-competitive assay systems using techniques such as Western blots, RJA. ELISA {enzyme linked imnu.mosorbcmf assay), 'Sandwich'' immunoassays. immunopreeipitation assays, immunodiffusion assays, agglutination assay % complement-fixation assay.¾.. immunuiadiomott'tc assays. fluorescent smmwmms&amp;ys* :atsd gratsiti A iraroimosssap.. Such assays am routine as d: wel ï known: in the au.

Antibodies can also bo assayed using any SPR-bawd assays known in the art for characterizing the kinetic parameters of the interaction of the antibody with t/5a. Any SRR instrument corm-nctxualiy available including, but not 1 routed to, BIAcore Instruments (Biacore AB; Uppsala, Swedenκ iAsys instruments {Affinity Sensors; Franklin. Massachusscttsk IBIS system {Windsor Scientific Limited; Berks. UK I SRR-CELLLA systems{Nippon Laser and Electronics Lab; Hokkaido, Japan», and SPR Detector Sprceta (Texas Instruments: Dallas. Texas) can he used in the methods described hereto, See, c.g.. Mullctt ct ai. (2000) Methods 221 77-4); Dong et a). (2002) AmAws4k 303-323.: Piwsh: ct ak {49983 &amp;0t:@pidMkipchnkif: 97 101. ar»d Rich et al (2000) Carr Opin Bioiechnol jjy 54-6 j.

It is understood that the above methods can also be used to determine if, e.g„ an anti-CSa antibody does not bind to full-length, native C5, C5, and/or (id proteins. The above methods can also be used to determine if an antibody that binds to C5u also inbibits the Interaction between CSa and a (. .5a receptor The above methods can also be used to detonmne if an antibody that binds to C5a also inhibits the activity of Ufa.

As described in the above references., after phage selection. She antibody coding regions from the phage can be isolated and used to generate \sholc antibodies, including human antibodies. or any desired fragments, and expressed in arty desired boat, including mammalian cells, insect cells, plant cells, yeast, and bacteria, c.g., as described in detail below. For example, techniques to reuombiuandy produce Fab, Fab and Fiat)"»;· fragments can also be employed using methods known in the art such as those disclosed in PCI publication no. WO 92/22324; Mullinax et ai, s 1902) Bioΐνι121(0:864-369; and Suwat ct ai. () 995s Am J R<pr Immunol ..)4.26--34, and Better· et ai (1988) Sdrncc 240:1(141-1043. Examples of techniques which can be used to produce singlechain Pw and antibodies include those described in U.S. parent nos 4,936,77s; and 5,258,498; Huston et al. 119° 1) Methods in Enz-ywoiogv 202:46-68: Shu et al. 11993) /Aw Naf 4<a,J Sr; USA 9(1/7995- /999: and Skerra et al |i9H8t Saom.i 240:1036-1040.

Phage display teehrieiogy can also he need to, c.g , increase the affinity of an antibody lor its cognate antigen. 'Hie technology, referred to as affinity maturation, can employ mutagenesis or CDR walking arid re-selection to identify antibodies thot bind with higher affinity to an antigen as compared to the initial or parents! antibody. See, c.g , Glaser etui. t lRR.c1,/ binuifi^l IdO: jfHG-T)ld .libraries start be constructed consisting of a pool of variant chines, each differing by one or more ammo acid substitutions. Mutant·»» with increased binding affinity for the antigen can be selected for by cementing the immobilized mutant1; with labeled antigen or any combination of methods described abuse. Ary screening method known in the art can be used to identify mutant antibodies with increased affinity to the antigen io,g.. SPR or ELlV\ exhmgacs}

In some embodiments, epitope mapping can be used to identify, c.g... the region of C5a that interacts v> ith an antibody, e.g., a region of C'Sa that binds to CSaR 1 Methods fot identifying the epitope to w hieh a particular antibody binds are also known in the an and arc described abuse.

The antibodies and imornents thereof identified hereir: can be or can be made "chimeric.’’ < hlmeric antibodies and antigen-binding fragments thereof comprise portions from two or more different species (c.g., mouse and humane Ohunertc antibodies can be produced with mouse variable regions of desired specificity fused to human constant domains to; example. V S Patent No. 4.816J67·. in this manner, nart-human antibodies can be tnodified to snake them more .suitable for human clinical application (c.g., methods for treating or pres enting a complement-mediated disorder m a subject}.

The monoclonal antibodies of the present disclosure include "humanized" fomw of the non-human tea·., mouse) antibodies. Humanized or CDR-«/.rafted mAbs are particularly usemi as therapeutic agents for humans because they are trot cleared from the circt.thuh.tn as rapidly as motive antibodies and do «rol typical;}' provoke an adverse immune reaction. Generally · a immunised antibody has one or more amino acid residues introduced Into it from a non-human source. These non-human amino acid residues am often referred to as' import" residues, which are typically taken from an "import" variable domain. Methods os preparing humanized antibodies are generally well known lit the art. for esample, humanization can be essentially performed follow ing the method of Winter 35x1 cs.'-workers {see, e.g.« Jones et al. (198M Sawn 32 ; '522-525; Rtcehrnann of ah 519sG) Nanny 332:323-327: and Verhoeven et al. {19K8) Si-fV«cr 239:1534-153bκ by substituting ’-«dent frameworks or CDR sequences for the corresponding sequences of a human antibody. AGo see, c g, Smelens et al. {200ό} .1/--,7 Gummo/ -13.-1243 - I 257. (si some emoodimems, human seed forms· of nom-h union ic.g.. mousej antibodies are human antibodies (recipient antibody) m which the CDR region amino acid residues of the nonhuman antibody (e.g., mouse rat, rabbis or non-human pnonite antibody * bin iog the desired specificity, Money. and binding capacity arc prat tod onto tiro IVamewotk -catb.dd of a human antibody. Additional h urn an mat ion methods are described below m the working extuopim.

Methods for grafting {.'DR sequences' from a donor antibody (e,g., a. eon-human antibody) to ihe framework regions of an accentor a mi body {e.g.. a hmmm autilxxb,·; are \>cH known in the art and are described in, e.g., Jones e? at. { Rhso) ΛG/ver 52 \ :522-525, 7- crhocyon et ai,(1988) 537.-,¾- 2j7(45-47): 15 34-;536: Rieehmunn et a: (1985) .Vu.Uf?c 552:523-32", Queen et ah {19Xy> Pn»: Mm Aom! $d VS.4 8b; 10029- i 0()55.: PC7 publication no. WO (!3'0 i 1 33". k.Mtkmo rough et ;-d. {{99') Prorth; dGM/ioermg. Aoum m/7 0' 4.7 AG7Mg Bonny K, G. Lo (2(8(4) "Antibody Fngineeviug- Methods and

Protocols," Humana Press {ISBN; !5k8290^2h; Borrcback (1992) "Antibody Engineering, A Practical Guided' W'.R, freeman ami Go. NY; and Borrcback {(905;. "Andbody Engineering," 2”" f.dmom Oxford University Press, NY, Os.ford. For example. CDI-G from a donor antibody can be grafted omn framework regions of an acceptor andbody using overlap extension polymerase chain reaction (PGR) icckmcpies as described in, e.g., Daugherty cf al. 1199; > Am/cA- .JmVG Pm 19(9):2-171-2470;

Rogtmka ci al. {R/%) Pr<m-.m Angóneouig GjOGMS-GMl; n-xl Yaanki et al. {2(8.(4) /7'me/ο e;g7mvrmg. Ao(go ά Sc/iV/hm s GoP-iS 1-959. in embodiments where tlic selected CDR. arnsno acid scigsenccs ace short sequence a te.g., tenor than SO- Id amino acids in length). nucleic acids encoding the CDRs i.an he ehenticaily syndics; red as described in, e g.. Shuukhi et al, {20b"·) \-<« Pa .dc.-Gs AY?,go? S' mm o'er ms 5 It 1): 129- igo and G.G, Patent No, 0,40^2 5 A por <5 green nucleic act'd sequence encoding an acceptor antibody, the region oCtho nucleic add sequence encoding the CDRs can be replaced with the chemically synthesized nucleic acids using standard molecular biology techniques The ,v and V ends of die chemically synthesized uucletc acids can he '.ynthcstzed to comtwisc sticky end restriction enzyme sites for use in cloning the nucleic acids into the* nucleic acid encoding the variable region of the doner antibody.

In sores; instances, one or more: fra;nee.oi'k region ammo acid residues of the human immunoglobulin arc also replaced by corresponding ammo acid residues of the non·· hut nan antibody (so cubed "'back nun,mens"), in addition, phage display libraries can be used to vary amino acids at chosen positions widun the antibody sequence The properties of a humanized and body are also affected by the choice of the human framework Furthermore., humanized and chimorized antibodies can be modified to comprise residue» that are not found in the recipient anti body or in the donor antibody In order to further improve antibody properties, such as, for example, affinity or effector function.

Fully human antibodies are also provided in the disclosure. The term “human antibody" me in dim antibodies having vans:.) in .and constant regions id present i domed from human immunoglobulin sequences, preferably human get mime sequences. Human amibodso- can include amino acid residues not encoded by human germune immunoglobulin sequences -c.g., munitions utmxlucen by random or sj re-specific mutagenesis in H/o: or by sontatie mutation in tvroi. However, the term "'human antibody" does not Include antibodies in which FT>R sequences domed from another mammah&amp;u species, such as a mouse, have boon grafted onto human framework sequences t'i.c., humunowd amlborirms, Fully human or human antibodies may he derived from transgenic ntice carrying human antibody genes (Currying the \ uriabie fV) diversity (D), joining fls, and constant |C) exonsi or from human cells. For example, it ;s now possible to produce hems genic animals (e.g„ mice·) tbut arc capable, upon imsnamzanon, of producing a full repertoire of human antibodies sn the so-cnee of endogenous immunoglobulin production See, o.g,, jblobos it» ei a I. i I hv3 s Pnh- opF Acad Sa IPS.i 10:2551.: iukobovit- ei al (Idv3t A'otun 16.TT55-.T5St Bruggemaun et a I \ ITto) }\·φ· in hwnnn>A. joy ami Bnchosai et si, i 1992) Ao/wv 555:25^. Tumsgems mouse strains can be engineered to contain gene sequences from unrearranged human immunoglobulin genes One example of sneb a mouse is the BnMAb Mmoese < Medarcx, Inc.), wind· contains human immunoglobulin uausgene miniloci that encode unreurranged human μ heavy and κ Hght chain immunoglobulin sequences, together with targeted manmens that Inactivate the endogenous u and sc chain loot. Sec, e.g„ Lonberg, of ah i loog) Satun· M&amp;i6474):856-859. The preparation and use of i luMab mice, and ihe genomic modifications carried by such mice. arc further described in Taylor os ai 19^2) Nucleic Add* Re< 20:6287-6295; Chen, .h cl ah 11993} International Immunology 5; 6-17-650; Tw.ohon c? al. {(993; /dm Xau uVoo St > (.-.5.-1 90 5720-37.74. Choi et al. (1993) \>itiire Gencfk\ 4,1 17. :23. Tuallion cf ad,; 1994)./ immuun! 15?:2!t 12-2920; Taylor ei al. « 1904) hiereeSve>ii /uom,u.s-do;;o 6:579--54) -raj π a he, ski -:-: ;.d. 11900} Yomsv Bionx he-i .1.4:845-851. An altorname transgenic monsc system Aar expressing human immunoglobulin genes is referred to us the Xcnornonse. (Ahgenlx, Inc.) and is described in, eg., U S patent sms. <\()75,161, ο.114.59Χ; 6,150,564; and 6,)62,9o3. Like tlte HttMAb Mouses system. the Xenomoust; system involves dea'upuon of the endogenous mouse heavy and light chain genes and insertion into tlte genome of the mouse mmsyeoes carrying imrcarrunged human heavy and light chain inrnmrioglobuiin loci that contain human variable and constant region sequences. Other systems known in the art for expressing human immunogjoliulin genes include the K..M MouseT-' system, described in detail in POT Publication WO 03/--13478 and the TO mouse system described' in Tot.meek a cf al (218)0} Proc Arid .AeaJ Sri USA 97.722-72?

The human sequences may code for hath the heavy and light chains of human antibodies and would function correctly in the 0:101% undergoing rearrangement to provide a wide antiixxl> repertoire similar to that In humanv Tlte transgenic mtee eat: be immunised with the target protein immunogen to create a diverse array tn specific antibodies and their encoding RXA bmcletc acids encoding the antibody chain components of such antibodies mas then be cloned irons the animat into a display sector Typically, .separate populations ui nudem acids encoding heavy and tight chair: sequences are cloned, and the separate populations then recombined on insertion into the vector, such that any given copy of the vector receives a random combination of a heavy and a light chain. The vector is designed to express antibody chains so that they cm be assembled and displayed on the outer surface of a display package containing the vector For example, antibody chains can be expressed as fusion protects with a phage coat protein from ?hc otuei sasiuce of ihe phage. Thereafter, d sa play packages can bo selected and screened for display of ask-bodies iiiodioe to a laren;, ]; aftdnithe nhnye-dNplay libraries screened above can include human amibodios (Hoooeuhoom et al (W92),/ Sfoj Biol 222:}$ 1. Marks et al. i i99!) ,/M»l BhB 222'58! - .va;; and Vaughan et al ( 1996} Λοη/ιν Bintcch 14:3091. bynthctk pftuye libraries cun be created o'hsck use randomized oosnbinafiorss of synthetic human antibody V-regions. B> ^cL-edon on andgan, huh human antibodies can be made in which the V-region-; are very human-dike m nature doe, e.y., U.S. Fluent Nos 6,704 ,132; 6,680,209, 4,634,660:. and Ostberg et al. s 1 984 ) Hx-hrUamu 2:341-2367, the contents of each of which art; incorporated herein by reference in their entirety. Γογ the generation of human antibodies also sec Mendez et at. (i 948;- \:(nurc (mmoles j.5' i 4m i do end Green and j;d<oU.>x,its ? 199b'} .> E\p Xk d i8b:-tk'3-49S, the disclosures of w such are hereby incorporated by so i or once sss their entirety. Human antibodies are further discussed and delineated in G S Patent Nos.: 5,034.598; 6,6/3,iJ86' 6,114,59b, 6,075.18 h 6J62,%3; 6,150,584, 6,715,6 K); and 6,637.103 a- vs ell as 13 S Patent Publication Nos. ::0030729905 A I. 20040010810.41, 200-10093622 M, 20000040363 Λ1,2(10500540.55 Λ1. 20050(196395 Λ1, and 200502s~o30 Λ1. See also International Publication Nos. WO ;)-Γ02ο02, WO 06-34006, and WO 48 24893. and Emopeau Patent No ftp (? 4o.3 151 B1, Use disclosure- of each of the ubove-ested patents, applications, and references are horens incorporated by reference so their entirety.

In an aUernale e approach, others, meknhng (ienpluoa· InternanonaL ha. have unload n "nmuloeus" approach, In the mm linens ..epprouch >m exogenous tg ίο·, ts is nd trucked de'otigh the inclusion of pm, os (indss uhud cones) irotn rise In lo;. u- Ί hits, mt*. or nunc \ n genes, one or mme bl; genes, one or more .0 genes, a mu eon-a.m- region, ansi a svsond constant region ipsvlesek-b, a gun η n a const tut region) arc ftvnvd into a construes for msenum tmo an anno,si this approach κ described m, ·.. g„ l .S. Patent Nos : 5,545,91) "; 5 545 80o, 5, ο,35,8.7 5; >,o25,G?,6. 5 f'33 i.75' 5,o6!,0So; 12Ί 5 ?b4,.p50, and 5,814.316. 6,5-o! ,Οο,ο; 5,o 12,205, 5.721 56". 5,780,2 j-y 5,013,^0), ö.èóo.spS; 5,.V"7,5o”; 309,129; 5,8'/ 1,790, 9,255,158, and ",Ο I 1,871. the disclosures of which are hereby mcotposxecd bv reiesessso m then enincu. See also kuropcun Patens

No, 0 546 073 BB hiiornauo.mil Patent Publication Nas, WO 92/03°! 8, WO 02/22645, WO °2 '2264/, WO 92/22670, WO 93-!! 2227, WO94 00569. WO 04 25585, WO 96.44450, WO 07/15653, and WC) OH'24884, the disclosures of each of which arc hereby incorporated by reference m their entirety. Sere further 'Taylor ct tit, 5 1992) Nuckk- . Ic/r/s i?rv 20: -0.287; Chen et ;d, 0 993) ha /nmumo/ 5: 647; TuaiUon et a! i 1994; proc Λ or/ Jcaif No/ I'SA 90: 4 720-4, Choi et al. -: 1994} Safurc GVmvrV.v 4: i 17; Lonberg ct al. 11994) ,\(i',c·,568; 856-859; 1 aylor et ah {1494; /oioevo.Omm //s-iumsv:,Oy > 0: 5/0--591; Fnadlon ct al {1995},/ dumms·'/ 154- 6453-05·; Ftshwild ct :4. (19961 Wno ίNrm/ogr jjc 8-15; and TeasΠοη a ah (2000) h}n):nn>>!. |0; 2949-4005, the disclosures of cosh of which «re hereby incorporated by reference m their entirety. hi certain enthodimeuK dc-lnvmuoired tones of the antibodies, or antigen-bi.ndi.ng fragments described herein an; pros idod, Dc-immurh.aed antibodies or a nopen-binding iragmemts thereof are ,mb bodies that huso been modified so as so render the antibody or a-mgen-binding fragment thereof Ui'Hwmmouogenit:. or less imntunogetuc, to d gisen species. Idc-iinmnnicndon cm; be achter cd by modi tying the antibody o: anneevi-binding fragment thereof utilising any of a variety of iucbifrqoex known so ihose skilled In the art (sec, e.g., PCX Publication Nos, WO (94 108!58 and WO 0094317;, For example, an antibody or anngcn-bistdmg fragment thereof teas he do-; noun rosed by identifying potential T ceil cpttopcs and/or B sad I epitopes within the amino acid sequence of the antibody or aobgcn-binding fragment thereof and removing one or more of the potential T cell epitopes and/or 13 cell epitopes from die antibody or antlgen-hiodiog fragment thereof! for example, using; recombinant techniques. The modified antihodx or amigen-binding fragment thereof may riten optionally be produced and tested to identify antibodies or amigcn-l.unding fragments thereof that have retained one or more desired biological activ itic-s, such as, for example, blinding aOl.nity, but have reduced immanogemcity. Methods -for identifying potential·"!" cell epitopes and/or B ceil epitopes, may be carried out using techniques know n it: the art. such as, for example, computational methods {sec e.g., PCT Publication No. WO 02-06924.2). //? vkro or 0/ s,>7/< <' techniques, and biological assays or physical methods (such as, for example, determination of the binding of peptides to MWC molecules, determination of the; binding of peptide:MHO complexe-; ίο the T cell receptors from (he species to receive the antibody or anmien-bindiny fragment thereof, resting of the protein ot peptide parrs thereof using rraasgertie animais with the MHC molecules of the species to receive the antibody or smigen-bmdmg freemen5 thereof, or lesiing u oh transgenic animal-reconsnuttcd ohh immune s\riem ceils from foe specie.-- to receive the antibody or etnioen-hinding i ragmen- rhereof, etc,}. in various emhoel intents. the dc-immuni/ed antibodies described herein include bc-irornuniaod antsgen-bmdmg fragments, Fab, io. seH, Fab' and F(ehfi;. monoclonal antibodies, murine antibodies. full) human anybodies. engineered anyhudi-m much as, lor example, ehnru rie. simde -, kun. ('DR-grafted. hiommmed, and a red i nel I y selected antibodies;, synthetic anybodies and semisynthetic antibodies.

In the therapeutic embodiments of the present disclosure, bispecific anybodies arc comemnbied. Bispecific antibodies are monoclonal, preferably human or .humanized, antibodies that have binding specificities lor at least too diHcrem antigens la the present case, one of Ihe binding specificities is for C5.k the other one is for any other antigen.

Methods for making bispecific antibodies are v- Ithtn the purs im,v of those chilled in the art. Traditionally, the recombinate prod tier teat of bispecific any bodies w based on the eo-espression of too immunoidobidln heuvv-cliain’lkdn-chain pah·.-, whom the o,vo hcsv\ chani light-chain pears har p different spceiiiciues 1 M doe in and Audio i 1 fix.¾i fidm/v ΜΑΤ.ΓΤΙνι. Antibods satiable domains oith the desired binding specificities {antHxrdy-andgencombining «itesi can be fused to immunoglobulin constant domain sequcttcès. The fusion of the heavy chum variable region i- preferably i- \>idt an immunoglobulin heavy-chain constant domain, inehtdutg at least part of the hinge, ( ;G. and Cud regions. DMAs encoding the immunoglobulin heavy-chum fusions and, if desired, the Immunoglobulin light chain, are inserted into separate expressions \eeiore, and an, eo-transieeted mm a suitable host organism. For further details of illu.sïmfise eurreolly knoon mcdmds tor gene on mg ίο-pom fie antibodies see. e.g., Surc-h ct ah ti9xo.1/- ihmls ,m d.’e.rr.ms Gmr _F?J/.:. 1 m PCI Publication l\o WO fib TTl 11. Brennan et al. 11fibp} SmVm e 229:8!. Shulabs et ah../ bb/.·· d/m/C 1992) |?$-217-225. Ko-tdny et a). (1 °*92J /a.>/??;).;;;>/ 148(3): 1347-155.1-: idol linger et ah i 199)) P? t.«. Xot! .Sum/ fi- / (PSA fillb444-o44H; Gruberet al. {199-4}./ immmm/ J55.5368, and Tuft ei al s tvfil;,/ hwtum»! I47-6Q, Rispees tie antibodies also include cro^s-i inked t>r hcicroconjugatc antibodies. I leteroeonjugatc antibodies may be made using any convenient cross-links ng methods. Suitable cross-linking agents arc well known in the art. and are disclosed in ITS. Patent No. 4,676.980. along with a number of ero.ssfoinking teehnicjuc&amp;

Various techniques tor making and isolating btspecific antibody fragments dirediy from recombined ceil euimre have also been described. For example, bispeedse antibodies tune been produced using, leucine dippers. See. e.g., Kostemy et al. (1992),/ ïmmtmoi 148(5}.-1547-1.553. The leucrae zipper peptides (tom tire Fos and itm pmtems may be linked to die Fab' portions of two di Herent antibodies by gene fusion. The antibody homodimers may bo reduced a? the hinge region to form monomers and them reoxidized to form the antibody heterodirners. This method can also be utilized for the production of antibody homodimers The '‘di&amp;body*' technology described by Hoilinger et al. (1993) Pror Nufi A-.aA Sci USA 90:6444-6448 has provided an aiternahve mechanism for making bi&amp;pccitlc antibody fragments. The fragments comprise a heavy-chain variable domain (VH) connected to a light-chain variable domain (VL) by a linker which is too short to allow pairing between the two domains on the same chain. .Accordingly, the VH and VL domains of one fragment are forced to pair with the complementary VL and VH domains of another fragment, thereby forming two antigen* binding sites. Another strategy for making bispccifk antibody fragments by the use of single-chain Fv (sePv) dimers has. also been reported, bee. o.gw Gruber ct at. (1994)./ ha/mino! Alternatively, the antibodies can be "linear antibodies" as described in, e.g.. Zapata et af {1995) PrvtUifi Eng. 8(1 QV 1057--1062. Briefly, these antibodies comprise a pair of tandem Fn segments iVu-Cu 1 -Vu-C’u 1} which tbrrn a. pair of antigen binding regions. Linear tuvbixxbos mm be bispecific or monospecific.

Antibodies with more than two valencies (e.g;.. trispeeifk antibodies} arc contemplated and described in, e.g.. Tint ct al. {199!} A Immune! .1.47:60.

The disclosure also embraces variant forms of multi-specific antibodies such as the dual variable domain immunoglobulin lDYD-!g} molecules described in \V« ct a.i (20(17} AOr Bivh'cknoi25( 11V. 12-)()-1297. The DVD-fg molecules are designed such that two different light chain variable domains {YL> trom two different parent antibodies are linked in tandem diteetlv or via a short im.ker by recombinant DMA techniques, followed by the light chairs constant domain Similarly, the heavy chain comprises two different heavy chain variable domains (VH) linhed in tandem, followed by the contant domain (";!.! and Pc region. Methods for making DYD~lg molecules from two narent antibodies arc further described in, e.g„ PCX Publication Nos WO 08/024188 and WO 07/0-4715.

The disclosure also provide* eamelid or dromedary antibodies (c.g., antibodies derived: from Cumelm Caieim dmMêéétM<% m imna pmcoii), Such antibodies, unlike the t> pical two-chain (fragment} or four-chain (whole antibody) antibodies from most mammals, generally lack light chains. Sec Ü S patent no. 5,'ëO.kbH, St o lemens et al. I :.00--0 J Bh>! Γ/u tn ΑΧ 1250- i 561, Dmnotdm et ;.d. (2003) h'iuan· 424;7X3-78b: and Plescltherger et si. (2003} Biorohja^tiie ('hum .14:440-448 Fnginecred libraries of varnchd antibodies and antibody fragments arc commercially available, for example, from Ably ns t Ghent. Belgium) An with other antibodies ofuon~ human origin, an ammo aetd sequence of a cam did antibody can be altered rveombinantiy to obtain a sequence that more closely resembles a human sequence, be,, ht; nanobody can be "humanized"1 to thereby further reduce the potential irntnunogcnieity of the antibody.

In some embodiments, the nnii-CvG antibodies described herein comprise an altered heavy chain constant region that has reduced tor no) effector function rerun e to its corresponding unaltered constant region, Effector· functions bn oh mg the consiaiu region of the asnn<'5a antibody may be modulated by altering properties of tire constant or Ft: region. Altered effector functions include for example, a modulation in one or more of the following activities: antibody-dependent cellular cytotoxicity {AIXXh, complement-dependent cytotoxicity (OX.), apoptosis, binding so one or more Fc-rccepiors, and pro-inflammatory responses. Modulation refers to an increase, decrease, or elimination of an effector lunet tost activity exhibited by a subject antibody containing an altered constant region as com pared to the activity of the unaltered form of the constant region, in particular embodiments, modulation includes situations in which an activity is abolished or completely absent.

An aherect constant region with altered feR binding affinity and/or ADCC activity and/or altered CDG activity is a polypeptide which has either an enhanced or diminished FeR binding activity aruTor ADCC activity and/or CDC activity compared to the unaltered form of ihe constant region. An altered constant region which displays increased binding to an FeR binds at least one FeR with greater affinity than the unaltered polypeptide. An altered constant region which displays decreased binding to an FeR binds at least one FeR with lower affinity than the unaltered lorrn of the constant region. Such variants which display decreased binding to an FeR may possess little or vto appreciable binding to tut FeR, e.g.., 0 to 50% to.g , less than 50. 49,4% 47. 4<\ 45. 44, 43. 42, 41.40, 39.. 3b, 37, MK 35. 34, 33.32, 3 i. 30, 29, 2X. 27. 20, 25. 24, 23. 22. 21,20, 19, lb, 17, 16, 15., 14, 13, 12, II. 10, % % 7. 0. 5. % 3, 2, or I %) of the binding to the

FeR as compared to the level of binding of a native sequence immunoglobulin constant or Fe region to the FeR Similarly, an altered constant region that displays modulated ADCC and/or CDC activity may exhibit Cither increased or reduced ADCC and/or CDC activity compared ίο the unaltered const am region. For example, ht some embodiments, the imf-CSa antibody comprising an altered constant region can exhibit approximately 0 to 50% (e.g. less than 50, -b>, :|8, 4?, 46, 45, 44. 43, 42. 4t. 40, 39.. 3H, 37, 36, 35, 34,33,. 32, 31, 39, 2% 28. 27, 26, 25, 24, 23, 22, 21. 20, 19, 18, 1?, im )5. 14, 1 5. 12, I R 10, 9, 8. 6, 5.4. 3, 2, or ; %) of the ADCC and or CDC activity of the unaltered Form of the constant region. An ant of’5a antibody described herein comprising an .altered cons tan! region displaying reduced ADCC and·or CPC may exhibit reduced or no ADCC' and/or CDC activity as exemplified herein ht certain embodiment'·., die altered uonMant region lies ai le no one annuo a on! substitution, insertion, anchor deletion, compared to a native sequence constant region or to the unaltered constant region, e g t'som about one to about one bundled ammo acid substitutions, insertions, and or deletions in a nanw seguc-me constant icgmn or in the con,stam region ui the n.-rent polypeptide In some embodiments, the altered constant region herein will possess at least about 70% homology tslnmang lor identity with the tmalten d constant region and in some instance." at least arvtu ”7% tnd m otlnu insumees at least about NO'*·.! homology or identity therewith and m other embodiments at least nbont 85°.!, 90% or °5% homology or identity iher».wnh The ahen.d constant region ntuy also contain one or more amino acid deletions or insertions. Additionally, the altered constant region may contain one or more amino acid substitutions, deletions, or insertions that results in altered posMranshnional modifications, including, for example. «π abetvd glycosyluiion pattern (c.g.. the addition of one or more sugar components. the löss of one or more sugar component;'. or a change in compositum of one or more sugar components relative u.> rhe unaltered consiam region).

Antibodies svfth altered or no effector functions may be generated by engineering or producing antibodies w tdt variant constant. Fe, or heavy chain regions; recombinant D'NA technology and Or cell culture and expression conditions may be used to produce antibodies with altered function and/or activity, For example, recombinant ON A technology may be used to engineer one or more amino acid substitutions. deletions, or insertions in regions {such as. for example, re or constant regional that affect antibody function including effectos functions. Alternatively, changes in post-uunslational modifications, such as. c.g., glycosylatiors patterns, may be achieved l.n manipulating the cell culture and expression conditions by which the antibody is produced. Suitable methods tor introducing one or more substitutions, additions, or deletions into an Fc region of an antibody are web known in the art and inducky e.g.. standard DMA mutagenesis techniques as described in. c.g., Sambrook et at. (1989,} ''Molecular Cloning. A Laboratory Manual, 2αά Edition,” Cold Spring Harbor Laboratory Press, Cold Spring Harbor. N.Y.; Harlow and Lane Π°ιχ). .<upru\ Bosreback. f 1999k sup's?; Johne et ah 119AH, supnr, PCT publication no. WO 06 Sent)); and (: S. patent no. 49)4,-1-41

In some embodiments, an ami-OSu antibody described hcretn exhibits reduced or no effector function. In so roe embodiments, an antl-CSa antibody comprises a hybrid constant region, or a portion thereof, such as a 111-(44 hybrid constant region {sec 0 g„ Burton ct at. (11*925 -/; A {mnum 5 1:1-1 A fan field et at. (199 h,/ Exp 3/W (Tv Ids A-1491; and Mue ller et al (19911) Mol dunnom/ 3-1(6):491 -452). See above. ht addition 10 using a G2 G4 construct as described above, an anti-CSa antibody described herein having reduced tdVeetor function mu> be produced by introducing other types of changes in the amino acid sequence of certain regions of the antibody. Such ammo aetd sequence changes include but are run limited to the Ala-Ala mutation described tit., e.g., PCI Publication nos WO 94-199)27 and WO 91--17531: and Xu et :4. (200ft) i'.’V/ !mniunt 4 200:1 o-2h. fires, in some embodiments, an a nu-( ha antibody with one or more ntutattons within the constant region ineluding the Ala-Ala munition has reduced or no eHector function According to these embodiments, the constant region of the antibody can comprise a substitution to an alanine at position 234 or a mutation to an alanine at position 233. Additionsi!y. the altered constant region may contain a double monition: a imitation vo an alanine at position 23-1 and a second imitation to an alanine at position 233. In one embodiment, ait ami-C5a antibody comprises an lgG4 frame work, wherein the Ala-Ala on;union would describe a rmuevioots; from phony Lb am no to alanuie at position 234 and or a nrutaiton born lencmc to alanine at position 235. In another embodiment, the umi-CSa antibody comprises an igG; framework, wherein ;he Ala-Ala rn;.itation would describe a mutatJott{st bora leucine to alanine at position 2114 ami or a mutation from leucine to aianme at position .:35. A.n ami-C5a antibody may alternatively or additionally earn, other rntnations. including the point mu union K.33.3 A in the (3112 domain (Monarch et al. <2001 ) J luv;/ J5A2 lob-121 eh}. At; antibody with said nuuationis t in the constant region may furthermore be a blocking or non-blocking antibody.

Additional substitutions that, when introduced into a heavy chair; constant region, rest»U hi decreased eifector function are set forth in, c.g, Shields et al. (2001 )> Βιοί i 'hi. pi 2 'Tab >;{S391 -6oQ4, bee panicularh Value I b'Btndtng of human IgG I variants to human Venn ami FeyR} of Shields et al. the disclosure of which is incorporated herein by reference in its entirety. By screening a library of ami-Ig.E antibodies, each antibody of the Uhary ditYenng lw one or mom substitutions in the heavy chain constant region, for binding lo a panel of be receptors i including FcRn. fcyRL FcyRIlA. RcyRIsB, and beyRIIIAK the authors identified a number of substitutions that modulate specific Fc-Fe reeenmr uncracuotis. Ida' example, a variant Igu2a heavy chain constant region in which the (Ί 12 domain contains a D2o5 \ so bob U: Lon (heavy chain ammo acid numbering according to K.abut ct al. i my oh·} results in a complete loss ofinteraotion bc sa veen the variant constant region end Ig(.l Fc receptors FcyR ίIB. FeyRlII, FcyRL and FcyRlV. Shields ct al, ί 20011 at page 0.345, Tabic I. See also Baudiuo et al. (2008)2 hiin^nau .1 .3:.1 .'0004-boo^ t \U7·.

Changes within the lunge region also atleet eilccto:' functions. For example, deletion of the hinge region may reduce a 0 miry for Fc receptors and may reduce complement activation (K.lem et al (1081) Prov Sad Acad USA ~H. 524-528}. The present disclosure therefore also relates to ami bod ms with alterations in the binge region. in «ι-if; embodiments, an ami-C5a antibody may contain an altered contain region exhibiting enhanced or reduced complement dependent cytotoxicity {CDC> Modulated CD€ activity may be achieved by introducing one or more amino acid substitutions, insertions, or deletions In an Fc region of the antibody. See. e.g., U.S patent no. 6,194,55 i. Alternatively or additionally, cysteine reslduds) may be introduced in the Fc region, thereby allowing uttcrehtun disulfide bond formation in tbi« region. The homed imerk antibody thus generated may have improved or reduced internalization capability and/or increased or decreased eontpicroont-rnedmrcd cell killing. See. e.g., (Von ct at. {1992),/ fixp Med 170:1191.|195 and Shopes 11992) Inmimol 14b:29IS-2922; PCT publication nos. WO 99/51692 and WO 94/2935 0 Duncan and Winter (1986) ANi/n/v 322:736-40. and U.S. Patent Nos. 5.648.260 and 5,924,821.

Another potential means of modulating effector function of antibodies includes changes in glycosylation, which ss summarized m, e.g., Rapa 1200.0 8k>Pwcf>.%· inrvnuuionul 1(4):44-53. According to Wright and Morrison, the mtcroheterogendty of human IgG oligosaccharides can atleet biological functions sue!; as (/(.')€ and ADCO. 'binding to carious be receptors, and binding to (Ίη protein. 099"? /7/2/70 7/,15:29-32. Glycosylation panerns of antibodies can differ depending on the producing cell and the cell culture conditions Raju, 0.-,000. Such differences can lead to changes in both; effector function and pharmacokinetics. See, e.g , Israel ct al i i 9%) .VmmVmv-87i,;i);575-57S; Ncnktrk cl al. < 1996; ( 79? Exp Bnmnvtl 10/>(2):239-2ρ4. DitTerens.es m effector function may be related to the IgCi's ability to bind to the Fey receptors (Feyks> on the oil ce is a cells Shields et al base shown that IgG, with alterations in ammo aenl sequence that have improved binding to FcyR. can exhibit up to 10(/6. enhanced WK ( using human effect»! cells. (2001)./ Biol Chow 2 /0(9) 9.591 -6604. Whiie these alterations include changes in amino acids not found at the binding interface, both the nature of the sugar component as well as its structural pattern may also contribute to the differences observed. In addition, the presence or absentee of fucose in die oligosaccharide component ot'an IgG can improve binding and AIX.'C. Sec. e.g , Shields ct ai. 12002).7 TO .7 (. O.-.w 277( 50).26?33-2o?40. An IgG that lacked a in cosy laten earl ;>( yd rale Itttkecl to Assr" exhibited normal receptor binding to the FcyR.I receptor. in contrast, binding to the FcyRIHA receptor was improved 5iMbki and accompanied by enhanced A DC , especially ur ion cr antibody concentrations

Shinkawn ci al. demcaisirased that an smithed} to the human ILo receptor produced trt a rat hybridoma showed more dien ATT higher ΛΙΧ C whets compared to the antibody produced us Chinese hamster oven, ceils tCHOj {Shhskawa et a! (2003»/Bi<7 Chan 2CM5)· Tibb-T.G. Monosaccharide composition and oligosaccharide piorïiing snowed that the rat hybridoma-produced IgG had a lower consent of i'acnse than site (flTO-produced protest?. The authors cone hided that site lack of fuoosv hUsem of an h;G 1 itu'i a critical role in enhancement ofAlX'C activity. A difTcrent approach na-.· taken by 1 ifuassn et a). w ho ehassgod the giycosylatson pattern of chCf:?. a chimeric IgG 1 anti-neuroblastoma antibody, (1 ;A)o; Am Buna /;?;<</ jJLü'i 7α·-ΙΚ(Π. Using vetracyeline, they regulated the aero sty of a giys.Oayknuisierase enzyme (GnTHD which Injects okgosaeehurhies that have been implicated in ADCC activity. Tisc ADCC act A isy of the parens antibody was» barely above background level. Measurement of ADCC activity of the ehCE? produced at different tetracycline levels showed an optimal range of fin nil evps'esskus tor maximal ehOIT ία c/ov ADCC activity This activity correlated with the level of covisiasst regson'associated, bisected complex oligosaccharide. Newly npinmzed variants exhibited ;»ubstamiai ADCC scsivny. Si sx si a sly. Wright and Morrison produced antibodies in a CHO cell line defies era in glycosy iation and showed that antibodies produced in this cell line were incapable of eornplemrmt-mediuied cyiolyals ί I WO»./ Eye .fCC 180' CC?- i 05>f». Thus, its known alterations that affect effector function include modifications in the glycos>lation pattern ora change tn the number of glycosylated residu·..··;, the present disclosure relates to an smu-Cea antibody whereto glvcosyiation is altered to either enhance or decrease effector tunedonts} including ADC 'ff and OOC. Altered ylycosyisHUot? includes a decrease or increase us the number of glycosylated residues as well us a change tn the pattern or location of glycosylated residues.

Still other approaches ον 1st for shoring she effector function of antibodies. Tor example·, amibody-producing cells oust be hypermutagenic, thereby generating antibodies w ish randomly altered polypeptide residues throughout ass entire asusbody nsolecule. See, e.g., PCX publication no. WO OTOi 1738. blyperruutagente host cells include cells deficient in DNA mismatch repair. Antibodies produced in this manner may be less antigenic andor have beneficial pharmacokinetic proporties. Additionally sneb amibodies may be .selected lor proporties such as enhanced or decreased ettccior fimctiontAy Add mental details tb' tnolectdar btology techniques use tui for preparing an antibody or amigonfoindi.ng fragment thereof described herein arc set forth below.

Reiombinans Antibody Expression and Purification

The antibodies or am’jgert-bmding fragments the? eot described herein can be produced using a variety of techniques known in the art of tnoleetuar biology and protein chemistry, for example, a nucleic aetd encoding one or both of rite heavy and light chain polypeptide» of an antibody can be inserted tmo an expression vector that contains transcriptional mu! translational regulatory sequences, which include, eg.. promoter sequences, dhosotna! binding sites, transeriptiesnai start and stop sequences, translational .start and stop sequences. transcription terminator signals, polyadonylnnon signals, and culuuieer oraui\ ator sequences. The regulatory,sequence's include a promoter and trnnsersphortal start and stop sequences. In addition, die expression vector can mclnde more than ot»e replication system such that it can be main coned in two different organisms, for ex-imple In mammalian or insect cells for expression and in a proiaryotie host for cloning cud amp!Uh.ation. bet etui possible sector systems are available for the expression of cloned heavy chain and light clean polypeptides from nucleic acids In mammalian celts. One class of vectors relics upon the integration ofohe desired gene sequences into the host coll genome. f el In winch have stably integrated ΡΝΛ can be selected by simultaneously introducing dang resistance genes such as £’. coh gpi t Mulligan and Berg 11 dx I } Pro·.. .Vo <7 ;.· aP Sc;' i 5.1 77:7077 t or 705 neo (Southern and Berg 1178,As \fo/A/yd C'.-act ,1:377). The selectable marker gene can be either linked to the DMA gene sequences to be expressed, or introduced into the same cell by co-transfection tWigjer et af {1770) s t-"/Z iff? 7 A second class- of vectors ufiltc.es ON A dentetnx winch confer auU'snomoush repikatiug capabilities to an extruchromosorna! plastnid These vectors can be derived from animal viruses, such as bovine papillomavirus (Sarvcr e; ;tl. t foKa) Pmc Wo'/ ,A.'t!;/ 5 a i '6,1, 7°;7147P cytosrtcgalov I rue, polyoma vints (Deans or al. (l^xd)

Pa>··: .Vo// Acad Sa USA 81; 1292), or SV40 virus (Lusty and Botchan (1981} Nawee 29327'».

The expression vectorscan be introduced intocells in a manner suitable for subsequent expression of the nucleic acid. The method of introduction is largely dictated by the targeted cell type, discussed below. Exemplary methods include CaPO,·, precipitation, liposome fusion, cationic liposomes. dcctropomtion. viral infection, dextmn-mediated transfection, polybrone-medialed transfection, protoplast fusion, and d i re ct m i c ro I n j c ct i ο n.

Appropriate host cells tor the expression or antibodies or antigen-binding fragments thereof include yeast, bacteria, insect, plant, and mammalian ceils. Of pare culm interest arc bacteria such as £. ctdh fungi such as Succharomyces eera-hiae and Pichia pa^toris* insect cells such as SI·'*), mammalian cell lines (c.g.. human cell lines), as well us primary cell tines.

In some embodiments, an antibody or fragment thereof can be expressed in, and punfied from, transgenic animals (e.g., transgenic mammals). for example, an atm body cun be produced in transgenic non-human mammals i'o.g., rodents) and isolated from nulk as described in, e g.. Houdcbine (2002) ('tor Opia Biou-chfn>/ .U{f.,j'i,2.>t>29; t an Kuik-Romeiiu ct ah (2000( Tmnsgatk· Res 9(2):)55-1588 and Pollock ct al. (1909)./ hamuad Methods 23j.pl;; 2j;. 1 J7-I57,

The antibodies and fragments thereof can be produced from the celts by culturing a bost cell transformed with the expression vector containing nucleic acid encoding the antibodies or fragments, under conditions, and for an amount of time, sufficient to allow expression of the proteins. Such conditions for protein expression will vary with the choice of the expression vector and the host cell., and will be easily ascertained by one skilled in the art through routine experimentation. For example, antibodies expressed in E. adi ear; be refolded from inclusion bodies (see, e.g.. Non ct al. {1998;· Cytokine lil» 19-30). Bacterial expression systems and methods for their use arc well know it in the cut <sce Current Protocols in Molecular Biology, Wiley &amp; Sons., and Molecular Cloning--A Laboratory Manual --3rd Ed., Cold bpring Harbor Laboratory Press, New York (2901)). The choice of codons, suitable expression vectors and suitable host ceils will vary depending on a .number of factors, and may be easily optimized as needed An antibody tot fragment thereof) described herein can be expressed m mammalian eel:* or in other expression systems including but not limned to yeast, baeukwsrus, and /?? r;Vo< expression systems {see, e.g... Raseabska et ah <20CKi> Protein Exprtsuon «»</ Purijiuiiio): fro ) S )--220). following expression, the antibodies and fragments thereto;' can be isolated. The term ‘'purified'' or '‘isolated’' as applied to any of the proteins {antibodies os' iragmetsist described herein refers to a polypeptide that has been separated or pun fled from components (e.g., proteins or other naturally -oewsn'ing luological ear organic moloeuieM which naturally accompany it, e.g., other pro terns, lipids, and nucleic add m a prokaryote expressing the proteins. Typically, a polypeptide is purified when it const;tutes at leas; kt) (e.g., at least 65. ?(h 75, kO, by, °0. 02, 95. 9'·. or ‘>9 ; ;fr, by weight, of the total protein ist a sfnnpie.

An antibody or fragment thereof can be s-ofried m purified it; a \ abets ut vwes known to those skilled in the an depending on what other components .no present in dte sample. Standard peril teal tost methods include eUxlt\>pUoreties molecular, immunological, and chromatographic techniques, including ion exchange, hydrophobic, affinity, and reverse-phase RfiLC chromatography For example, an antibody cat be purified using a standard anti-a mi body column (e g.., a protein-A or proteinO column) UUrafihraiion and diafuirauon techniques, in coni unction wish protein concentration, arc also useful. See, e.g., Scopes (1994) ‘Protein Purification, 3"f edition/' horinger-Veriag, New Vork C sty, New·· dork The degree of purification necessary will vary depending on the desired use. In some instances, no purification of the expressed antibody or fragments thereof will be necessary.

Methods tor determining the yield or purity of .a purified antibody or fragment thereof are known in the art and Include, e.g., Bradford assay, liV spectroscopy. Biuret protetn assay, ί.ον,τν proiein assay, anode black protein -assay, hig.it pressure liquid chromatography tiiPLC), mass spectrometry {MS), and gd electrophoretic methods te g , using a prevent stain such as C'oonnmsie Blue or colloidal slker stein t.

In some embodiments, endotoxin can be removed from the antibodies or fragments Methods for removing endotoxin from a protetn sample arc know n hi the art and exemplified in the working examples. For example, endotoxin can be remov ed from ft protein -simple -using a vancry of co-nmereiaiiy available reagents including. without limitation, the ProtooSpin5 :'· Endotoxin Removn! Kits (Nonnen Biotek Corporation»,

Detox i-C.l el Endotoxin Removal Gel (Thornto Scientific: Pieren Protein Research Products), MiraCLFAN u; Endotoxin Removal Kst (Mims). or AciCichsoïv - Mustang-'^’ P mem bra ït o (Pa i I ( 'orporut ion). .Methods tbr delecting and.'or measuring tb o amount of endotoxin present m a sample (both betere and after ρηηϋοηοοη) .are known in the art and convener;.!,>; kits are available, Foe example, dte concentration of endotoxin ist a protest'; sample can be determined using the QCl.- i 000 Chnomogtmc kit (Bio Whittaker), the lunulas mnebocyte lysate (LA!..Phased kits .such as the Pyeoteil'x·, PyrotelbR-T. Fyrockrome-x. f'hromo-Ι.ΛΕ, and CSE hbs aren I able from the Associates of Cape Cod Incorporated.

While ht no nay intended in be limiting, exemplary methods for generating the antibodies described herein are sc* torth in the working Examples. ΜίΜΒΑΕΡ.Ι.ΙΕΟ..Β!ΐ.ύ; .si .A n.t i bodtex m A m.1 gcn rBhtdmg Pragma ilia amiboTes or antigen-binding fragments thereof can be modified follow mg their expression and purification. The modification.-» can be cot aleot or non-cosaieot modifications. Such modifications can be introduced into the antibodies or fragment·- by. e.g , reacting targeted amino acid residues of the polypeptide w itk an orgaine den vat mete agent that is capable of reacting with selected side chains or tot initial residues (suitable sites (or modification cart be chosen using any of a variety of criteria including, e g , structural analysis or ammo acid sequerwe analysis of the antibodies or fragments.

In some embodiments, the antibodies or antigen-binding, fragments thereof can bo conjugated ?o a heterologous moiety. The heterologous motet) can be, e.g.. a heterologous polypeptide, a therapeutic agent (e.g,. a toxin or a drtsgh ora detectable label such as, hut not lime red to. a radso.ae.itve label, an enzymatic label, a fluorescent label, a heavy metal label, a luminescent Uthel, or ait affinity tag such as bsotin or streptavidin. Suitable heterologous polypeptides include, e.g.., an antigenic tag to g., Ei.AG (DYKDDDDK -SEC) ID ΝΟΤΟ}), pol) histidine (MJk It HI I Hid 11 ;SFQ ID N():K|), hemagglutinin (HA: YPVDVPDYA (SFQ ID NOTc}», glutathlonc-S-iratisfera.se (GST), or maltose-binding protein i MBP»» for use m purifying the suit bodies or fragments. Heterologous polypepndes aho include polypeptides (o g.., cmrymes) that arc useful as diagnostic or detectable marker'*, ibr example, Ineifemse. s fluorescent protein ic.g,, green fluorescent protein (Gi:P>h or ehloounphemcol acetyl fmnslLrase (CAT) Suitable mdloscthe tubch include, c.g.; "f. -'IT ’ V. ;:Ί. : ‘Ί. *‘S. and ΊΤ Suitable fluorescent labels include, without limitation. Ituoresocivt, fluorescein isoihiucvauafe ;ΓΠC'}, green fluorescent pooien· fGFPk Dy LightrM 4SS, phycoerythrin (PEL pmpMlnm iodide (PH, PerOlL PH--Aiexu Fmo<m· 700, ('yd ,tlioph>eoeyunm, and Cy7. Luminescent label·- include, e.g., any oi a r ariety of luminescent kmtksmdc te.g., europium or terbium) ehchnes. For example, suitable europium chelates include the europium chelate of diothylenc triamlne pentaacem: acid (O'ΓΡΑ } or t e r r at x s c y e 10 d 0 d eca u e L A 7, iO-te;rauceue acid i f.K>TA). Enzymatic labels include, e.g., alkaline phosphatase, CAT, lucifcrase. and horseradish peroxidase.

Two proteins (e.g., art antibody arid a heterologous moiety) can be cross-linked using any of a number of known chemical cross linkers Examples of such cross linkers an; those which link to0 amino acid residues via a linkage that includes a "lundered" disulfide bond. In these linkages, a disulfide bond a Ithm the crons-linking uno Is protected Toy hindering groups on either side of the disulfide bond) from reduction by the action, for example, of reduced glutathione or the enzyme disc hide reductase. One suitable reagent, 4~suectni?rtidyiosycarhovi} j-i^-methyhu(2-pyridYldithto) toluene (SMPTb forms such a linkage between two proteins utilizing a terminal hsinc on one of the proteins and a terminal cysteine on the other. Hctcrobifuncuunai reagents that crosslink by a ddTerem coupling moiety on each protein can a ho be used. Other useful -crosslinkers include, without limitation, reagems 0Inch hnk i00 amino group*» (e.g., N-5-azido-2-n irrobenzoyloxysuccinireide). too sulfhydryl groups (e.g.. M-bts·-maldmiaobunrucL an tuniuo group and a sulfhydryl group (c g., m - m a I c i rn; do b c st c o y L χΙ -hydroxy-sueeinrencle ester), an amino group and a carboxyl group (e.g.. 4--[p-azidosalicytamidojlmtykunine;, and an amino group and a guantdnuurn group that is present in the side chain of arginine te.g,, p-exidophcnyl glyoxal monohydratc),

In some embodiments, a radioactive label can be directly conjugated to the amino acid backbone of the arm body. Alternatively, the radioseme label can be included as past of a larger molecule (e.g , '"'Ί in meta~(1 ‘ 'T{sodophonyi'N~hydm\ysuecimmide ij ‘‘"'I jt'nlPNHS) which binds to tree armno groups ίο lorn? rncot-iodophens i irrbPl derivmn es nuclei am pro terns (see. o g... Rogers ,;t a!, I9P7) J Wb .1 k:api;; 122; -12203. or chela :c (e <.... vo DOT Λ or DTP 4} which is in turn hound ίο the rooie in backbone. Methods of conjugating the radioactive labels or kir ger molecules chelates containing them to .he anuhodics or anbn.cnhanding fragments dewni>ed herein are known in die art. Such methods involve incubating the proteins e nd the radioaoux c label under conditions (e.g,. pH, sad conecnirabon, and or tempen.mu\o that iacdn.ne binding of the radioacnse label or chekue ίο the proiem (see, e.g . U b. Patent No 6,001,52^).

Methods fof conjugating u Ouoreseem label (sometimes referred to as a Mnsorophore'd ιο a provem ie.g,, an antibody) arc know n In the art of protein chemistry Pot example. fbaorophores can be eonunyued ίο bee amnio groups (o g., of lysines? or sulrhydrs 1 groups (e.g., cystetnes) ol proteins using succinirmck 1 (Nillfx) ester or letratluorophenyl (IT P) ester moieties an ached to the bnorophorcs hi sonic embodiments, ihe fluorophores cun be conjugated to a hetcrobifunctional cross-· tinker moieis such as •mild-SMbT'. Suitable conjugation methods msolsc incubating an antibody protein. or fragment ?hereon with the Ouoropbutv under eotuhtions that facilitate binding of the fluorophore to the protein. Sec, e.g , V\ cleh at;d Redvaoly (2005? "Handbook of lladioplianoaecmieHls' Radtoehemtstrv and Applications," John Wtky and Son" nSBN 0-3 71 405Ó05 i.

In sonic embodiments, tlic antibodies or fragments can be modified, e g., with a moiety that improves the stabilization and Mr retention of the antibodies in etrenkuton e g , in blood, serum, or caber tissues, for example, the antibody or fragment can he PRO) luted as described in. e.g.. Lee οι al i P-IROj /Gw,mjuy Chan 10(6): 0"'e-ft kt ostler et ?il. (.:002 3 Urufs /.!ν.η·ιν··"ηΌ Pcvtirn.s pd .4 7-4-5.7: and Roberts ei at. io002) AJvatuvJ /deni; ik‘ih\'ty hVWcns b-i :--150-4 eo or HJbSykifed (btvsenius Kahn Germans: see, c g., Paristc et al. (201 {3) hi*./ Pharm 1187(1:.2).: 1 VO-110) The stabilization moiety-etui impiorc the stability, ot retention of, the antibody (or fragmentR-, at lea-u 1 5 ic- g , at leas? 2, 5. UK 15, 20, .75, 50, -10. ur 50 or more) fold. hi some embodiment». die antibodies or antigen-binding (Vagrneois thereof described herein can be glycosylated .In some embodiments, an antibody or antigenbinding hrugrncro thereof described herein can be subjected to eneyrnabc or chenueal treatment, or produced from a cell, .such ihai die antibody or bugment has reduced or absent glvcosyhuon. Methods lor producing antibodies with reduced glycosylaiion are known jo die an and described in. e.g., U.S. patent no. 6,9.3.3,368; Wright c: al {iTol) £MBOJ JillJiD·-7 i 7-27231 and Co ct ai. (!9«3> 3/-,-/ hnmunot 30- * 361. fobU'.i'^

Compositions containing an antibody or an antigen-binding fragment thereof described herein can be formulated as a pharmaceutical composition. c.g , for administration to a .subject for the treur ment or prevention of a complement-associated disorder The pharmaceutical compositions will generally include a pharmaceutically acceptable carrier. As used herein., a '‘pharmaceutically acceptable carrier'" refers to. and includes, any and ail solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and die hke that are physiologically compatible. The compositions can Include a phurmuceusaaiiy acceptable salt, e.g.. an acid addition salt or a base addition sail - see,, e.g.. Berge ei al. {IPhanv. Sd 66. Md).

The conipositions can be Ibrnitilated ncwording to standard methods. Pharmaceutical formulation is a vvcll-esiablished am and is further described m. e.g., Gennaro {,7.000) “Remington: The Science and Practice of Pharmacy.'' elf*1 Edition, Uppineott. Williams &amp; WiH-.ins t iSBN. 0683306472); Ansel cl al, ί 199“) ".Pharmaceutical Dosage forms and Drug Delivery Systems.'’' TK Edition. Lippincou Williams &amp; Wilkins Publishers (ISBN; 008)305777); and Kibbe (3000) “Handbook of Pharmaceutical Excipients American Pharmaceutical Association? 3:d Edition t ISBN: 09I733096X). In some embodiments, a composition can be formulated, for example, as a buffered solution at a suitable concentration and suitable for storage at. 3-foC {e.g,, 4% 3 In some embodiments, a composition cart be formulated for storage at a temperature below- O H? (e.g., -ZiY'C or -80^0. In some embodtroenfs. the composition cam be formukued for storage for up to 7 years (e.g , one month, two months, three months, font months, five months, mx months, seven months, eight months, nine months, 10 months. 11 months, I year. .1G years, or .7 years? at 7-8°C fo g.. 4 (7). Dins, in some embodiments, the compositions described herein ere stab!·,; in storage for ut least I year at 2-H' C te.g., 4":Ci.

The pharmaceutical compositions can he in a variety of tbrrtH. These forms include, e.e„ liquid, scon-soud and solid dosage hums. such as liquid solution·; io g„ injectable ami in I visible solutions). dispersions or suspension··., mblets, phis, ponders, liposomes and suppositories Ί he preferred form depends m park oi; the intended mode of administration and theraprtdie application, For e earn pie compositions cennunme an anobody or fray·nee; intended Fm «.ysienitt or mmd ddwery ran be m the Farm ot tnicctublc or infusible sol·.·dons. Accordingly she compositions can he formulated for administration h\ a parenteral mode ie g . iorrav,-irons subcutaneous. nurapvrimneG. or moamuscular injection t " Pure u tern! adnmmemnoof 'domes:cred pan. moral v, ' and other gramimumady equivalent phrases. as used herein, referte modes of edmuususuum other than enteral and topical abmunsnatioo usually by injection, and include, Without hnmahon, intravenous tntranasai, intraocular, immmuvulir. Intraarterial mtrathce.d. nmocapsoku.. uunuirbital, mo .icardiae. fiUmdermal, intrape;uoneaJ, transtracheal, subcutaneous, subcuticular, inrra.tittcuh.tr, snbv.apss.ibr, subarachnoid, immspmal epidural intracerebral, insrac; smal uinacarohd and smmsternal injection and udissioo.

The compositions can he formulated as a solution, microomnlmm, dispersion, liposome, or other ordered structure suitable ibr statue storage ut high concentration. Sterile injectable solutions can be prepared In incorporating an ant;bods (or a fragment of the antibody) described heroin tn the required amount in ast appropriate solvent with one or a combination of mmedlools cm; me; ated above, as required, followed by filtered stalli?.;nloo. Generally, dispersions ate prepared by Incorporating an antibody or 1 ragedescribed herein into a sterile vehicle that contains a bash dispersion mcdmtn and the ieqm red other ingredients from tho.m enumerated above. In the case of sterile powders ibr the preparation of sterile Inject able solutions, methods tor preparation include vacuum drying and iree/c-drying that yield a powder of an antibody, or an antigen-binding Tragtneui thereof, described herein phis any additional deseed ingredient.· (see below-1 from a previously stcrde-dUcred solution thereof The proper fluidity of a solution can be mammined, for exampl.-. by the use of a coming such us lecithin, by the maintenance of the required particle size in the ease of dispersion and by t he use of surfactants, Prolonged absorption of injectable corn position- can lx; brought about by including so. foe composmon a reagent that deug s ikoorphoru for example rvsouosteanne salto, and gelatin

The aott-< 5a antibodies. or antigen-binding fragments thereof, described herein can also be formulated in immunoliposomc compositions Liposomes cr.nti.ai.ning rite antibody can be prepared by methods known io the art sod; as. c.g., the methods described in Epstein et al. t.iv8Sj Prov Kat/Mad Sci US-1 *2:3688;. Hwang et al. (N80) Pmc NaP AcdP Sd USA 7?:4()30-. and U.S. Patent Nos. 4,485.045 and 4,544.545. Liposomes w ith enhanced cocuiunon time are disclosed in, e.g., U.S, Patent No. 5.013.55.6.

In ccroon enilxxImiotsN. an enubods or an antigen-binding fragment thereof -..art be prepared with a carrier that o til protect the compound agaotsi rapid release. snelt as a controlled release fonrmhntott, including implants and microencapsulated delivery s%stems. Biodegradable, biountgxnibk· polymers can be used, such as ethylene vinyl acetate, polyanhydndo-% polsgiscolic at.id, collagen, polyoohocstets. and polytaUtc acid. Many methods for the preparation of such formulations arc known in the art.. See, e.g., 3.R. Robinson "'Sustained and Controlled Release Drug Del is cry Systems.''

Marcel Dekker Inc , New York

In -<ome embodiments, an antibody or antigen-bindmg bagmenr described herein can be ft t$ initialed in a composition suitable lor Intrapubnonars administration tc.g,, for administration Ua nebulizer, sec belros ) to a mammal such as a human. Methods for preparing such compositions ure o mi known in the art and described in. c.g;., U.S. patent application publication no. 70040307513: U.S, patent nos. 7,112,341 and 6.0 i 4,%b; and PUT application publication nos. WO 00 001 !~b and WO Do 12.2257, fee disclosures of each of which are incorporated herein by reference in their entirety.. Dry powder inhaler formulations and suitable systems lot administration of the tbrmuknious arc described in, e g., 1: S patent application publication no. 200702^5029. PUT Publication Nr;. WO 00 OWxT and U.S, patent no, 536)7,544,

In some embodiments, an anii-<'5a mm body or anttgen-binding fragment ibereot described herein can be forntithned in a composition suitable tor delivery to the eye In suntc embodiments, one or more of the anü-Cda antibodies (or antigen-binding fragment.-thereof) described bereist aas be admensfowd locally, tbr example, by way of topical application or intraviircai injection. For example, in some embodiments, the aoU-C'5u. antibodies can be formulated (or admhtixtxatfon by way of an eye drop.

The therapeutic preparation for treating tbe eye can contain one or more of the ;mti~C5a antibodies in a concentration from about 0.01 to about 1‘w by weight, preferably from about 0 05 to about 0.50¾ in a pharmaceutically acceptable solution, suspension or ointment, the preparation obi preferably be in the form of a sterile aqueous solution containing, e.g , additional ingredients sneb as, but not it mi fed to, preservatives, buffers, tonicity agents, antioxidants and siahmuers. nontontc wetting or e fori tying agents, and v 1 scosi ty- meres sing agents

Suitable preservative', for uve m snelt a solution include hcrmalkomum chloride, beneethonhtm chloride, chiorobnumol, thimerosai and the like. Suitable buffers include, e g., boric acid, sodium, a mi potassium bicarbonate. vtxlntm and potassium borates, sodium and potassium carbonate, sodmm acetate, and sodium biphospluno, it? amounts sufficient to maintain the pH at between about pH 6 and pH M and ptelcrahly, between about pH 7 and pH 7.5. Mumble tonicity agents are desman 40, dextran 70, dextrose, glycerin, potassium chloride, proneterte gtyeol. artd -odium ehlortde.

Suitable amiwnduMs and stabs beets include sodium bisulfite, sodium rnoiablaiiinte. m nil urn thiosultue, and thiourea. Suitable wetting and dertfytng agents utclude potysorbatc hi), polysorbate 20. poloxamet 282 and tyioxapol. Suitable viscosity increasing agents .include dextmu Mb dcxtrtm M.b gelatin, glycerin. hydroxyethyleethdosc, Hydrovsnetlsylpropylcelluhtse, tanohm methy iecliulose, petrolatum, polyethylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, and cnutxsxymcihyiecliulose. The preparation can be udmimstetctl topically to the eye of the subject sr; need of treatment (e.g , a subject at the ted with by com cmhmnS methods, e.g.. in the form of drops, or by bathing the eye in a therapeutic solution, containing one or more nrüi-Cêu antibodies.

In addition, u variety of devices have been developed for introducing drugs into the vitree! cavity of the eye, for example., H.S. patent application publication no 200200201 7b describes a phamutceuUeai-contuiuing plug dial cart be inserted through the sclera such that it projects into the vitreous cavity to deliver the pharmaceutical agent into the vitreous cavity, in another example, U.S. patent no. 5,-1-13.505 describes an implantable device ior introduction into n suprachorotdal space or an avascular region for so ruined release of chug into tin; interior of the eye. IAS. parem no.v A7AA ON and 6.00 s 3X6 each disclose an hnnlsntiihie drug delivery device attachable to the scleral surface of an esc. The device comprises an inner core containing an effective amount of a low solubility agent covered by a non-bioerodibie polymer that A permeable 10 the low solooitity agent. Darnig operation, the low solubility agent permeates the bioerodible polymer cover tor sustained release out of the de% ice. Additional methods mid devices to g.. a transscicm; patch and delivery via contact lenses) for deliver)·· of it therapeutic agent to the eye are described in, c g.. Ambari and Adamis «-002» (Vug /for/.» Eye Rvs Hlfo'l-fo' fob. Kama and l =rt u {A) 06} A/e Drue fl/mri Ah s >Ai.j.A Ho IA fo I. Btuoeas and Batadtandran lAOOb i AA/vrr Opw Drug De/nvn At I i: I I Of 10 ;·. Ofoken and rhauhan foOlH s A vest Of^huimoi ΓΑ S 4 4AA'd A A447; K im ot ;d. i.?00“) Oyhihaimn AVs JAAMA!54, and PC"!' publication no. WO 04 0705 A the disclosures of tv inch an. ineorpoiated herein by reference In their entirety.

Nucleic acids encoding an antibody tor an antigen-binding fragment thereof) cats bo incorporated into a gene cons weet to be used as a part ce a gem, therapy protocol to deliver nucleic acids that can be used to express and product: «gems vs nbin cells isee below t Fbxpression constructs c>l such components may iv admnfoaered so; any therapeutically effective carrier, c.g.. any formulation or coniposiifon capable of etïCi.H>el\ deux urine the component gene to cell·' ;>? ;yo.>. Approaches include insertion of the subject gene in viral vectors including recombinant retroviruses, adenovirus, adeno-assoeiuted virus, Antivirus, and herpes simplex vnus-l tHSV-1 i. or recombinant bactcrm I or eukaryotic plasmids. Viral vectors can trans leu cells dhvouy: plasmid DN A can be delivered vvith the help of, for us ample, cationic liposomes s Itpoiecfln) or d on v a hoed ie.g., antibody conjugated), polyiysmc conjugates, gramicidin S. arimfoul viral en velopes or other such mfrateiluku· earners, as oeH as direct in jection of the gene construct or CAiPO,; precipitation tsce, e.g„ WOCM 06(1-4)4) earristd out in v;Ao Sec also, "A.v rfvn Approaches." below.) hx am pi eh of suitable retroviruses include pl.,3, pZ IP. pWF. and pf M which are know η to those skilled m the art (see. egg, P.glitis et al. (I'"foe) S nr?;;v AA): IAIAA ifob; Danes and Mulligan t b-A8i Prth - Sad Acad Sri i 5 -1 .bA.o-Ibb-6464, Wilson ot ah t l°bfo /Von Sad A<.ud Sc; l.'S.i 85:4014-4018; Armcniuno et al

Ν&amp;ίί, Aetië, 'USA USA 88:8039-8045; Ferry et ai. (1991 \ Froc Nat! Ac*u! Sci USA 88;83?7-8381; Choxvdbury et «I. (1991} Science .7.34:1802-1895: van Beuseohem ei ai {1992; Proc Nat! Ac cd Sci USA 8fj;7t>40-?644; fiay et al. (1992) Human Gene Thempy 3:64}-947; Dm ei al 11992} Pree Nat! Acad Sci i \\4 89:10892-1089$: Hxvu et ui. (1493),/ fnitnufuA 150:4104-4115; ü.S. Patent Nos 4.868, i 16 and 4,980,286» PCX Publication Nos WO89/07136, \V089 Π2468. WÜ89/053-I5» and Wi>92.07573). Amuhor omi gene delivery system indices adenovirus-derived vectors Dcc, e.g.. Bcrkner ei al (19881 8ioftcht;Aiiics 6:616; Roseoteld ct al. f 1991} Scion;? 252:431-434; and Rosevdeld ei al. {1902; G.7/ óK: 143' I 55}. Suitable adenoviral vectors derived front the adenox irus strain Ad type 5 did 24 or other strains of adenovirus (e.g,.; Ad 2, ..343. Ad?, etc.) are know it to those skilled In the art. Yet another viral vector system useful for delivery of the subject gene is the adeno-associated vitas (A AY). See, e.g.. Rotte et al. 11902} Am,/ Rc^n'r CNI ,J/o/S7o/1:3-19-356: Samulski et a). {198-4)./ 17m/6^:3822-3828; and Mcl.aughlm et al. i 1989),/ iW 62 }9o3-!973.

In some embodiments, an antibody, or amigen-bmdmg fragment thereof, described herein cart be formulated with one or mote additional active agents useful for treating or preventing a complement-associated disorder in a subject. .Additional agents for treating a complement-associated disorder in a subject will vary depending on the panicu[at disorder being treated, but can include, without limitation, an amihypertensixc (eg., an angiotensin-comertingenrvmc inhibitor;.. an anticoagulant, a corticosteroid ie.g , prednisone), or an immunosuppressive agent te.g., xinertsune or cyclosporine 3x1. imam pies oi anticoagulants include, e.g., warfarin tCoumudm'). heparin, phenindtone, tondapacinux, tdrupunnux, and thrombin inhibitors ie.g.< uigah'oban, iepirudin. buabrndin, or dabiganan) .An antibody or fragment thereof described herein can also be formulated xx itl; a fibrinolytic agent ie.g , srtermi. e-nminoeaproic acid, anlipjasrnse~a;. prostacyclin, and deftL'totitic.) lor the neat ment of a complement-mediated disorder. In scene embodiments, an tmnlxviy can be formulated with a lipid-lowering agent such as an inhibitor of hydrosymethylgkaaryl CoA reductase In sot no embodiments. an antibody can be formtilated with, or for use with, an atut-i.T>20 agent such as ntuxltnah < RhusaGG Btogen Idee, Cambridge, M.\). In some embodiments. e.g , for the treatment of RA, she antibody or rmtigen-lunding th-gment theseoS van \k formulated with one or both of infliximab {Rcoueadei'. { 'entocor Inc ! and methotrexate {Rheumatrex-'t·'', Trexalfob In :-0 no: e nfood ernenR an a no. body or :ni am:gca~bmdfog fragment thereof described herein can be formulated with a non-steroidal ;mufo:Hnmrn,itory dreg iNSAlD). Mans different NSAIDls are available. some mci the counter including ibuprofen (Advil an, Mot; h;a!', Nuprin es and naproxen (Ahevo-Kg avid many others arc axallifole by prescription including mcloxienm (Muh;RM, etodoUte (Lodmeafo, mfournetone {Rdaieaeu. snlindac {i'hnonR ), tfoomemm fl okv un-rfo, choline magnesium so 1 icy late «Trihisute^o, diclofenac {ifosaflRA, Voltaren-A Aifotrotecfofo DtHusinas (Dolobsd'”1), in do metmesafoalocio-Αί, Keroprofen fOrudlsX. OrnvaiMp, oxaprocm i Pay pro A:, and pmr-uearn t Feldenc-ny In some embodimems an antibody or a fragment thereof can be formulated for use oh if an anti-hypertensixe, an imis-seizurc agent te g., magnesium sulfate), or an aon-tinomhobe agent, bnti-bypertensixes induik, c.g., labetaloL hydralazine, nifedipine, calcium channel antagonists, nitroglycerin, or sodium nitroprusshie. See, e.g., Mihu et ah {20()7)./ Gasroinunitt lh-cr Dis. lp(d):d 19« -12-1. Ami-thrombotic agents include, e.g., heparin, andrhromhrn, ptostacyclhvor low do-e aspirin

In some embodiments, an antibody or antigen-binding fragment thereof can be formulated for administration to a subject along with intras enous gamma globulin therapy tIVIGh plasmapheresis, or plasma exchange. In some embodiments, an ami-C5a antibody or antigen-binding fragment thereof cart be formulated for use betere, during, or after, a kidney tmnspUnu.

When an antibody or antigen -binding fragment thereof is to be used in combination with a second active agent, the agents can be formulated separately or together, for example, the respectixe pharmaceutical compositions can be mixed, e.g.. just prsor to administration, and administered together or can be administered separately, e.g., at the same or different times (see below).

As described above, a composition can be formulated sue It that tt Includes a therapeutically effective amount of an :mrt-C.5a anhbody or antigen-binding fragment thereof described herein. In so;no embodiments, a composition can bo formulated to include a sub-therapeutic amount of the sub bo ay tor fragment) and a sub-ihctapcuric amount of one or more additional active agents such drat the components m total are thera pern ready effective for treating or preventing a complement-associated disorder. Methods for determining a therapeutically effectiv e dose oi an agent such as a therapeutic antibody are know?? in the art and described herein.

AneMoim:- J’he antibodies, antigen-binding fragments thereof, <.onjugatcs. and compositions of any of the forego trig can be used m a number ot diagnostic and therapenbo applications, for example, detecu.tbty-labeled anu-Cba antibodies te.g... anti-human 05a antibodies or anti-mouse 05a antibodies; can be used in assays to detect the presence or amount of 05a present in a biological sample. Determining the amount ol €5a m a. sample, e.g. a patient blood sample, can be useful to evaluate the lose! of complement a cm anon in the meurde. Suitable methods for using the antibodies in diagnostic assays are known in the nr? and include, without limitation, FUS.5. -moresccnec resonance energy transfer applications. Western blot, and dot bio? tcchmcptea. Sec, u.g , Sambrook et a 1., end Auxubel et sl.-xupm. in some embodiments, the antibodies «nd a»ftgcn~bindittg fragments described harem can be used as positive controls m assays designed to identify additional novel compounds for treating comp I ement-modiat-ed disorders For example, an aniOC5a antibody that Inhibits €5a activity can be used as a positive control m an assay to identify additional compounds (e.g... small rnolcculca, aptaroers. or antibodies? that inhibit €5a or OSa-denendeut C5a receptor signaling.

In some embodiments, the cross-reactive anh-Cea antibodies or antsgen-btndtng imgmunis thereof (mg , cross-; eactivc with human 05a and, e g., eynornolgu-s macaque C5a? desejlbed herein can be used for nre-cbnicai testing in non-human mammals, e.g,. pharmacokinetic or pharmacodynamic Studies in non-hum tv; primates. Accordingly, a researcher Wishing to evaluate the efficacy, of an ami-05¾ anti bods in treading u complemem-smsoeiaie disorder of interest iv.g,, R.A or sepsis) can use a cross-rcaciK e anti-05a nmibody described herein tn an appropriate non-human primate model of the disease. .If-be researcher, for example, establishes efficacy of the antibody in the no inhuman primate model, these results may provide sufficient nroof-oi-coucept support tor regulatory approval for use of she antibody in treating humans. Alternatively. or in addition, uresearcher may administer the uce>ss-rcueti\e antibody to a non-human primate u> study, e a., antibody clearance and or phnriWieodyuwrhcs properties. Based on such studies using the eross--reaeti\ e antibody, the researcher east bettor approximate the dose requited to treat human disease, irt some embodiments, the ami-mouse 05a antibodies or antigen-binding fragments thereof described herein, as well as antibodies that crossroad with lotman and nrou-sC C.'5a. can be used as ,a sttrrogatu antibody in mouse models of human disease. This can he especially useful w here a i tenet meed umi-human CM a amt body does not crossreset with mouse Cent anchor is likely to cause an anti-human antibody response in a mouse to 'aided the humanized antibody Is administered. Accordingly, a researcher wishing to study the effect of tut umi-Cöa antibody in beating a disease (e.g. ischemia-reperfusion injury) can use an amt-mouse ( 5a antibody described herein in an appropriate mouse model of the disease. If the researcher can establish efficacy In themes use model of disease using the unit-mouse C?a antibody, the results may establish proof-tti-concept tor use of an anti-human (da antibody in treating tite disease m hurnans. "The working examples disclose ats exemplary study using an anti-·nouse C5a antibody surrogate in &amp; mouse model of RA establishing proof-of-eoncept tor the use of an ami-human 05 a antibody to treat RA in man.

The umi-lMa ami homes described herein eat; also be used in methods for purifying C5a from a sample (e.g., a biological sample) In some embodiments, an tmit-t Ms andbody can be t turn obi lies'll on a sol Id phase support using methods well known m the art. Λ sample containing the antigen to he purified, in this case C5a, is contacted to the antibody on the -.olid support under conditions and for a time mfikaem to allow the antigen to bind to the mas body. The solid support is then washed one or more times with a .suitable buffer to remove unbound material The solid support can he then contacted with a second buffer that results in the release of the antigen from the antibody. The released antigen h then collected and elnaoc termed fe.g,, for purity and acne it\ ? using we'd known methods in the an.

The ami-CSa ami bodies and amigew-bmdmg fragments thereof described herein can also be used in therapeutic methods as elaborated ou he low.

The a!x.»vc-dehcHbcd compositions are useful in. ?>//<.'*· «·///< o methods for treating or preventing a variety of complement-associated disorders in a subject. The compositions can be administered to a subject., e g.. a human subject, using a vnriets of methods that depend., in part. on the ronre of administration. Tbc -cm te can be. e.g.. mire venous injection or infusion ti v'b cuheumomms injection fSO, mtrajerntonom (ΙΡι injection, of irmtamusetdar injection tl’Ms.

Adamustmtion can be aethered by. cap. local infusion, injection. or by means of an implant. The implant can be of a pen ms, π cm-porous, or gelatinous material, including membranes, snelt as mala a be membranes, or fibers. The implant can be configured for „vu stained or periodic release of the composition to the subject See. e.o., IJ.S. Pa tent Application PnKhcaiion No. 200b{.C41 Tim ITS. Patent Nets. 5.501.KÖ6; 4.1441,45 ?, attd 5 A10.795; f.P4SS40i; and f.P -130539. the disclosures ot each of which are incorporated hereto by reference tn v'neu entirety. The ttomposthor; cart be delivered to the subject by way of an implantable device bused on. e.g.. diffusive, credible, or eonveUn,e s>stems, e.g , osmotic pumps, biodegradable implants, elect rod if in si on sememe, eleerrottstnosfs systu-tTim vapor pressure pumps, eleetrolytie pumps, offers escem pumps. pie.-coeleeb'ic pumps, comm m baaed systettrs. or electromechanical systems.

In some earned!meats, an mm-Cda atuibuds oratnigen-binding fragment thereof h therapeutically delivered to a subject bs way of local administration. As used herein, 'local adrnoiisttiition'' or "local dcirt cturefers :o delivery that docs -to! rely upon transport of the composition or agent to its intended target tissue or site via the vystuhn system for example, the composition may be delivered hr. injection or implantation of the composition or agent or b> injection or implantation of a c!e\ ice containing the composition or agent, folio ving local administration in the vicinity of a target tissue or she, the composition or a gem. or one or more components thereof, may diffuse to the intended target tissue or site.

In some embodiments, an unti-Cêa anti hods or antigen-binding frag roem thereof can be locally administered to a joint te.g„ an articulated jouuj. for example, m embodiments whore the complement-associated diamder is arthritis, dut complement inhibitor can be admunstered dl reedy to a joint 'ey.·.· uno a joint apices or m the \ ieinhy of a joint. f. camples of mttaarueuhr johus 10 which an a no-0:-.: antibody or ants gen -bi.ndi.np kapmesit thereof can be locally administered include, e.g.. the bin. tome, elbow, wrist. sternoclavicular, tentperoinandibuhtr, eatpal, tarsal, ankle. and any other joint at;b|ect to arthritic conditions. An antbCea antibody or ..mdgmebwdlng fragment thereof can aho be administered to bursa cue it as, c.g., acromial, btcipitomdtak cuhitoradtak deltoid, infrapatellar, ischial, and any other burse known in the art of medicine. in some embodiments, an anti-Cpa anbboch or nmigcn-biudiug fragment thereof can be locally administered to the eye. As used herein, the term 'eye" refers to any and ail anatomical tissues and structures associated with an eye. The eye hew a wall composed of three dobed las ers, the outer sclera, the middle choroid layer, and the inner retina. The chamber behind the leus is filled with a gelatinous Huid re (erred to us the vitreous humor. At die back of the eye ?s the retina, w kick detects light. The cornea is ,m optically transparent tissue, which conveys images to the buck of the eye. The cornea includes one pathway for the permeation of drugs into the eye. Oil ter anatomical tissue structures ussocituud with the. eye include the lacrimal drainage system, which includes a secretory system, a distributive system and an excretory system The secretory system comprises w motors that arc stimulated by burking and temperature change duo. to tern evaporation and relies secrctors that have an efferent parasympathetic nerve supply and secrete tears In response to physical or emoUona! stimulation. The distributive system includes the eyelids and the tear meniscus around the lid edges of an open eye, which spread tears over the ocular surface by blinking, thus reduetitg dry areas from dev eloping.

In some embodiments, an uml-Cën antibody or antigen-binding fragment thereof is udmimstesed to the posterior chamber of the eye. In some cirkaxlimems. art ami-OSa antibody or antigen-binding fragment thereof is administered intra vitreaity. In some embodiment*, an andA. xt antibody or antigen-binding fragment thereof is administered trans-scleraily.

In some, embodiments, e.g , in embodiments for treatment or prevention ot a complement-associated pulmonary disorder such as COPD or asthma, an ami-C'5a amiktdy or antigen-binding fragment thereof described herein can also be administered to a subject by way of the lung Pulmonary dreg delivery may be achieved by inhalation. and administration by inhalation hetvtn may be oval und'or nasal Examples of pharmaceutical devices lor pulmonary delivery include metered dose inhalers, dry ponder inhalers DPls), and nebulizers l or example an anti-Ova antibody or an antigen-binding fragment thereof can be administered to tire lungs of a subject Ivy w ay of a dry powder inhaler. These inhalers are propellant-free dev iets that delis or dispersible and stable dry powder formulations to tbs- lungs. Dry powder inhalers are well known t.u die an. of nic-JIt me and include, w idiom Imthmlon: the TurboHaforU’ t AstraZeneca; I. ondort, England.! the AIR·*" inbaier I Alkev;ru..sA, i.ambndge, MassachusettsV, Rotabalenr: tfdlavoSnuihivllnet London. England); and Eellpw:ïi; (SmuTi-Anonus: Paris, France!, Set also e.w, PC 1 Publication Nos. \\ O 0-1.0.NafSO, WO 04 0.7.4 I ;;6, and \\ O 01.'7X0*EE DPI dev ices bas e been used for pulmonary administration of poly peptides snub as hisuhn and growth honnone. In some onthodlntents, an antE-('5.a antibody or an ainigen-binding fragment thereof can be iofnipulmonanly administered by way of a metered dose Inhaler. These inhalers rely on a propellant to deliver n discrete dose of a compound to the lungs. Esarnpios of corn pounds administered by metered dose inhalers include, wg., Asiovenrw iBoehringer-ingeiheirn; Ridgefield, f'ormecdcut 1 and Fins ent-w (GlasoSt'nithK.tirtci. See also, c.g..; U.S. Patent New. 6,170,717; 5,-14 7,150, and 6,005,141 r

In .'onto emhodmtomx, art anti-C5u antibody oranttgen-btnding fragment thereof can be administered to the lungs of a subject by way of a nebulizer. Nebulizers use compressed air to deliver a compound as a hqneffod aerosol or rnist A nebulizer can be, e g., a jet nebulizer ic,g., air or liquid-jet nebulizer?) or an ultrasonic nebulizer.

Additional devices and imrapulmonary administration methods are set forth in, mg., U.S, Patent Application Publication Nos. 70050771660 and 200001 tdn/'O. the disclosures of each of which are incorporated herein b> reference in their entirety.

In some embodiments, the antibodies or antigen-binding fragments thereof provided herein are present tn unit dosage lorow winch can be particularly .suitable for seiforaministrahon, A formulated product of the present disclosure can be included w ithm a container, typically, for example, a ved, cartridge, prcfillcri syringe or disposable pen Λ doser such as the doser device described in EES. Patent No. 6,70.7,855 may also be vised, for example, with an injection system of the present disclosure

An injection system of the present disclosure may employ a delivery per; es described In ü.S. Pk are No. 5.308.34;. Pen devices. mo.sj commonly need lo;· wei-deüvery of insulin to patients with diabetes., arc well know n in the art. Such devices can comprise at leest one injection needle {e.g.. a 3} gauge needle ol about 5 to n mm m length ¢, are typically preduled with one or more therapeutic uni; doses of a therapeutic solution, and are useful for rapidly delivering the solution to a subject wtMt as little, pact as possd-k

Ota. medication delivery pen includes a vut I holder into which a vial of insulin or other medication rnay he received, The v ial holder is an elongate generally tubular structure with proximal and distal ends The distal end of the vial holder includes mounting, means for engaging a double-ended needle cannula. The proximal cud also mcludes mounting means for engaging a pen body which includes a driver and dose setting apparatus. A disposable medication Keg., a high concentration solution of an ami-· 05a anti body or imtigen-bmnhig fragment thereof) containing \ ial for use v, ith the prior art stal holde; includes a distal end having a pkrcenbic elastomeric *ep?urn that can be pierced by one end of a double- ended needle cannula ' The pros mud end of this vial includes a stopper slidably disposed in fkud tight engagement with the cylindrical wall of the vial. This medication delivery pen is used by inserting the vial of medication into the vial holder. A pen body then is connected to the proximal end of the vial holder. The pen laxly includes a dose setting apparatus for designating a dose of medication to be delivered by the pen and a driving apparatus for raging the stopper of the vial d is tally for a distance corresponding to the selected dose. The user oft he pen mounts a double-ended needle cannula to the distal end of the vial holder such that the proximal point of the needle cannula pierces the septum on the v ml. The pat unit then selects a dose and operates the pen to tttgc the stopper distally ίο deliver the selected dose, f he dose selecting uppumtus returns to ;mro upon injection of the selected dose The patient then removes and discards the needle cannula, and keeps the medication delivery pen m a couveniem location for the next required medication administration. The medication in the vial will become whacked after several Mich administrations of medication. The patient' then separates the vial holder from the pen body. The empty vial may tkts be removed anti discarded. Λ new vial can be insetted tnto the vial holder, and the vial hinder and pen body can he reassembled end used as explained above. Accordtstgh, a medication do) A cry pen generally has a dme mechanism for accurate dosing and ease of usd. Λ dosage mechanism suds as a rotatable knob a!Soo s foe user to aoeumtciv adjust the amoatu of medication that will be injected by the pen from a prepackaged via! of medication. To inject die dose of medication, the use? inserts die needle under the skin and depresses die knob once as far as it o il! depress. The non may he an entirely mechanical device or it may he combined n idi electronic circuitry to accurately set and or indicate the dosage of medication that is injected into the user. See U.S. Paten;

No. 6, tt-r.Th'di. in so site embodiments, the needle of the pen device is disposable and the kus include one or more disposable replacement needles. Pen dev ices suitable tor delivery of the any one of the presently featured antibodies or antigen-bmdnig fragments thereof arc also described in, e.g., U.S. potent nos, 6377.003 of.KhoU’-hv. and i>, i =16361, the disclosures of each o r which are iticorporated herein by refercrice in their entirety, Λ os rerouted le-bused pen device is described in. e.g , U.S. patent no. 7350,615, the disclosure of which is incorporated herein by reference in Us entirety. Sec also the Precision Pen injector (PP!) device. Mo333 maun inau red by Scandinavian Health Ltd.

The -present disclosure also presents control led'-rcloase or extended-release formulations* suitable for chrome and or sctUadrnntistration of a medication -me!; as an ami-C5a antibody oran antigen-binding fragment thereof described herein. The various formulations can be administered to a patient in need of treatment \> sth the medication as a bolus or In continuous infusion -over a period of time.

In some embodiments, ,-¾ high concentration xnft-C 5a antibody for ttufigen-liinding frag met tf thereof! described herein Is formula red tor sustained-release, csrended-release, timed-release, cont-oUed-rclease, or continuous-release administration. In some etnbodiments, depot rormnkdions are used to administer the antibody to the sub-ket in need thereof. In this method, the antibody is formulated with one or more earners providing a gradual release of active agent over a 'period of a number of hours or days, Such tonnulaiious am of Urn based upon a degrading marris a inch gradually dispenses in die body to release the active agent hi some embodiments, a C5a-bindiny fragment to. si... 3 single chain inn; body, a dtabody, or a Fab' fragments of an asm-{.’5a andbody described herein is administered by way of mtrapuhnonary adothusmiiion ίο a subject is; need thereof. For example. a .single ehasn am shod y form rsf any of the anii-Cda antibodies described herein can be delivered by may of a nebulizer or an inhaler to a subject {e.g.. a hot nan) afflicted with a complement-associated puimonurs disorder seek as asthma or COPP. Λ suitable dose of an antibody or fragment thereof described herein, ahich dose is capable of treating or preventing a cotnpientertt-aasoeiatcd disorder tn a subject, can depend on a variety of factors including, cm. the age, sex, and weight of a subject to he treated and rite particular inhibitor compound used For example, a different dose of a ά hole utiti~C5a antibody may be required to treat a subject with R h as compared to the dose of a C?a-binding Fab' antibody fragment required to treat the same suhieer. Other factors a Hoc one the dose administered to the .subject include. e g,. the type or severity of the complometu-niediated disorder, for example, a subject having R.A may require administration of a different dosage of an imti-t.'Sa antibody than a subject with AMD. Other factors can include, e.g.. other medical disorders concurrently or previously affecting the subject, the genera] health oft I a subject, the genetic disposition of the subject, diet, time of administration, rate of excretion, drug combination. end any other additional therapeutics that arc administered to the subject, it should also he understood tied a specific dosage and treatment regimen for any particular subject will also depend upon the judgment of the treating medical practitioner f e g., doctor or nurse)

An antibody described herein can be administered as a fixed dose, or nt a milligram per kilogram img;kg} dose. In some embodiments, the dose can also be chose·! to reduce or asoid production of antibodies or other host immune responses agamst one or more of the active antibodies in the composition. While in no way Intended to be !uniting, exemplary dox«g> of an antibody, such as an anti-CSa antibody include. e.g„, I -1000 ug.'kg, 1 100 ug-'kg, 0.5-bO gg kg, 0,1 100 mg·kg, 0.505 ugkg, i -10 ng-kg. and I-10 itg kg. I-100 mg. kg, 0,5-50 rng kg, 0.1 -100 mg. kg: 0.5-55 mg-'kg, 1-20 mg: kg. 0,100 mg kg ro 1 mg-kg, and I -10 mg'kg. Exemplary dosages ol an antibody or antigenbinding fragment thereof described herein include, without limitation, 0.1 ugkg.

Cl;S 1$ gg%g> 2.Θ 'pg/kg,.4 agffeg,: smd: % 1¾¾¾ ö. 1 ffig/lg, 0J llö mg/lsgf 2.(5 mg/kg, 4 rng/kg. 8 rng/kg, and 20 mg/kg. Λ. pharmaceutical composition can include a therapeutically effective amount of an anti"C5a antibody or aniigenOinding trag-nont thereof desonhed herein Such effective amounts can be readiU determined by one of ordinary ski it in iite art based, in pan, on the effect of the administered antibody, or the combinamnid erfcct of the antibody attd erne or mote additional active agents, if mote than one agent is used. Λ therapeutically effective amount of an antibody or fragment thereof described herein can also vary according to factors such as the disease state, ago. .son, and weight of the individual, and the ability of the antibody (and one or more additional active agents) to ebr.it a aestieci response in the individual e.g., amelioration of at teas? one condition pacamemr, e.g., amelioration of at least one symptotn of the complement-mediated disorder. For example, a therapeutically efteehve amount of an antbf pa antibody can inhihu (lessen the seventy of or eliminate the occurrence of? and. or present a particular disorder, ard-'or any one of the symptoms of the particular disorder known in the art or described herein. Λ thentpatticitily effeettve amount is atso one in which arty toxic or detrimental effects oi the composition are outweighed by the therapeutically beneficial effects.

Suitable human doses of any of the antibodies or fragments thereof described herent can further be evaluated in. e.g.. Phase l dose escalation studies See, e.g., van Gufp et al. (2QÖK) Am J Transplantation 8((0:1711-17 IK; Hurtouska c? al. (200"!) Cih;

Gweer Rvs .ng2,..part.jj:52 3-531.; and Hetherirtgton et al. (2006} Atitwurrohial A gams' ami 50( 10): 3499-350Ö, 1 he terms mhetapentiealK offer lise amount''m mhempeut lenity efioctive dose," or similar terms used herein are imended to menu an ammtni of an agent (e.g , an ants-C5a antibody or an antigen-binding fragment, thereof) that will elicit the desired biological or medical response {e.g.. an improvement in one or more symptoms of a complement'- assoc sated disorder). In some embodiments, a composition desert bed herein eon mins a dicrapcuncaliy effective amount of an antibody, or antigen-binding fragment thereof, which specifically hinds to a neo-epitope present m C5a. in some embodiments, the composition contains any of the antibodies or sntlgcn-hindtng fragments thereof described hero ia and one ui more {c.g., (m\ throe, four, five, s;x< seven, eight, tune, !0. or 1 i or more) additional therapeutic agens such tha; the composition os o «-hole ss therapeutically cfk. et we. lor example, a composition cat: co nu o o an ;mti~C5a antibody demri heel herein and on rninumosupptosxix e au unie wherein the antibody anti agent are tacit at a coocenO'aüon that when combined are therapeutically effective for treating err [Meverging a e o m ρ Ï e me o t -- a s see s a t ed disorder ie.g., a oornpierncnt-asaociated inilammatmy disorder such an f'OPD, asthma, scorns, or ΚΛ) in a snhiece

Toxicity and therapeutic efficacy of such compositions cm; be determined by known pharmaceutical procedures in cell cultures or expert mental urn maïs te,g., animal models of any of the complement-mediated disorders described herein). Use of an ami-<;5a amtbody m an am mat mode; of RA is exemplified in the working examples. These procedures can bo used, e g.. for determining the !,XA> {the dose lethal to 50Ά of the population) and the ED-(! tine dost dtempeuttcadv affreuse in 50‘T of the population). The dose ratio luetweert toxic and therapeutic efiects is the therapeutic index and it can be expressed as the ratio Ι..ΙΧ:. RTfo;. An antibody or antigen-binding fragment dier cot" that exhibit-s a blah therapeutic index is preferred. While compositions that exhibit toxic side effects may be used, care should be taken to design a delivery -xsiotr; that targets such compounds to the site of affected tissue and to minimize potential damage to normal cells and, thereby, reduce side effects

The data obtained from the cud endure assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such antibodies or aniigen-bmding fragments thereof hes generally wuhin a range of circulating concentrations of the antibodies or fragments that include the bfW> v oh π til·,.· or no toxicity. The dosage may vary within tins utugc depending upon the do-age form employed and the route of administration utilized, for an umi-05a antibody desen bed hereoi, die therapeutically effective dose can be estimated initially bom cell -culture assays. Λ do.se can be formulated in an mud models to udbexe a circulating plasma concent mi ion range feat indudes the HTdi.e.. the eonec-urration of the antibody which achieses a half-maxima! inhibition of symptoms) as determined m cell culture. Snelt information can be used to more nee mutely determine useful closes ;u humans. Levels in plasma may be measured, for example, by high performance liquid chromatography. In some embodiments, ο g., ό here iocs I adnsmirirarion (c.g., to the eye or a joint) ta doored, celt m-bure or animal modeling cun be used so determine a dose required to achieve a therapeutically e it eenre concentration within the local site in some embodiments. the me diode can he performed in conjunction nidi other therapies for complement-associated disorders. For example, the composition can be administered to a subject at the aarstc time. prior to. or after, plasmapheresis, IVIG therapy, or plasma exchange, bee. e.g., Appel et al 0005? J Am So( VtyAral lo: 13921404. In some embodiments, the composition can be administered to a subject at the same time, prior to. or niter, a kidney transplant. A '\ahi.ct ' m used Iscwm, can !v any mammal hm' example, .a -mJm.et ran t\ a hum in t non-human primate to e - moo key, baboon, or cinmpanreot. a horse,:i cow, a rig. a sheep, .·, uoat a doe, a ear, u tabba, a guinea pw, a gerbsf a ha mat or. a rat. ora mouw. in some embodiments, the sub-tee: ta an infant te g., a human doanO.

As used herein, a .subject Os need of prevention.' 'm need oi tieatnvntf or “in need dieseo)/' rotbrs to one, who In, the judgmoui of .m .umfopnat,, nkdn.a] praeuiioner 5 e.g., a doctor, a nurse, or a nurse practitioner in the ease of humans; .¾ -, otermanan in the case of non-human snansmids?, would meson ably benefit bom a giver; trearmem (.such as treatment with a cosopcmooo comprising an anth-Cfe. antibody)

The term "preventing" is art-recognUed, and when used in relation to a condition, is vmh understood in the art, and me holes administration of,; corn position which reduces tlte frequency of, or delays the onset of, symptoms of a medical condition in a subject relative to a subject which docs nm reecive the composition. Thus, prevention of a complement-associated disorder such as asthma includes, for example, reducing the extent m frequency of cough mg., wheeamg, or cheat pain in a population oi patterns receiving a prophylactic treatment relative roan untreated control population, and or delaying the occurrence of coughing or wheeling in a treated population versus an untreated control population, e g , by a statistically and or clinically significant amount.

As described above, the antibodies and bfoiodeaMy-uetivc fragments described herein can be used to treat a variety of completncni-assoeiatcd disorders such as, but nn? limited to: rheumatoid atlhmis i RA ?; lupus nephritis, tsehemiii-reperfnxlon injury; atypical hemolytic uremic syndrome (aftUS?, typical or infectious hemolytic tsremte syndrome plH ‘S>. dense deposU disease iPDI)>, nuumsmal riociuntnl kcmegkd>ir>ur<,i >;PSUk mitmptc '-.doros5 > (MS?, mactdm du generation te.g., age-reLued mueuku degeneration i.\MDit: hemolysis, cletnued IKer enzymes and lov% platelets ij ik 1.1.Pt ndivono; sepsis, denmuontyosius; chalvdu retmopathy, dtromhonc thr-umboey mpemu purpura {TTP>. spontaneous fekü hms; IVnci-immune \ avcalnis. epiderm-dysis bullosa, ret'iim-ïii ie oh lom; muhtpic sderosts {MS); and imumunc brain nnury Sas, c.e. , I loleis «-UOHi mernssmss.m:·. m; AS>a n '·. 443.300-a 1 o and Ikders and I hunrmn i4004; Gohmem /mme/mé/gr 41; l-P- 1 52 hï lome embodtsricrjm. the <..o;nptemvtst~meds<ues1 die;· nier ^ a eorrspiomem-mediated s ascuim dismdet such a,s, hat nos bran cd na, a cnrd;outsudar disorder, myocarditis, n cerehrm,asrdar disorder a peripheral ie y,. irnmcnmskehrut; i > aseuUu disorder, a rono\ asenkn disorder, a menoueuo eutenc t ascurn' disorder, rosaseulurieunmt 10 ïranspb-,η?·- and or replants, s aseukiix, Uenoeh-Sckonkm purpura nephntts, -y sienuc mpus esy titernatosa,·- -ns^ociuied \ .mcuinis, v aemdmx associated o ah rheumatoid snhraG. immune complex \ Usubem, ! aker, nsu s disease, caphte.ry leak syndrome, dilated enidirurivoputhy, dodende anyiopaihs, ihorasie-abdornniHi sortie aneurism, Kanvusaki's disease iancrdls}, venous pas enihohis (VGEj, and restenosis ibliomlng svem placement rotational atherectomy, and percutaneous non is 1 urns red coronary angioplasty {PTCΛ r {Sec, c.g„ U.ö. patent application publication no. 40070; 7,;4S3 1 ba some embodiments, the co 01 p S o η nv π i - assoc ia ted disorder is myasthenia goo is, co kl - a ggUtt min disease tC'AD), paroxysmal cold hemoglobinuria (PCH), dennaiomyositis, scleroderma. v>,arm autoimmune hemolytic anemia. Graves' disease, Hashimoio’s rhysoiduix, type I diabetes, psoriasis, pemphigus, autoimmune hemolytic anemia (ΑΙΗ.Λ), idiopathic thrombocytopenic purpura i ΓΓΡ?, Goodpasture syudrotnc, «nbphosphoiipid syndrome t APS), Dcgos disease, and catastrophic APS (CAPS). hi some embodiments, ;m anti-(35 a anhbody or atnigcod.bridiog. fragment thereof described herein, alone or in combination wait a second anti-inflammatory agent, can he used to treat an in Hamms lory disorder such ns, bur not limited to, RA (ahose), mthmoKnoiy bowel disease, sepsis {above5, septic shock, acute lurtg injury, disseminated intravascular coagulation {.DIC.h or Crohn's disease. In some embodiments, the second ami-inti ammatory agent can he one selected from the group consisting of NS AIDs, corticosteroids, melhotre.xah;, hydroxy cbloroquinc, anU-TNF agents such as ctanerccpf and Hdlixmieb, a B coil depiction agent such as rituximab, an intetleuktn-l antagonist, or a T cell eostirntdarory blocking agent such as abatacent.

In some embodiments, the complement-associated disorder is u complement-associated tmurological disorder such as, but not limited to, amyotrophic lateral sclerosis fALSs. bruin injury, Alzheimer's disease, and chronic mOmmoatony denn- o I inuring neuropathy.. ( ompiemcnt-associalod disorders also Ineiede cturnplcmenoausoi.tumd pulmonary disorders such as. but not linnred to, asthma, bronchitis, a chronic obstructive pulmonary disease (CUPDi, an interstitial lung disease, o>S anti-trypsin deficieoo}, emphysema, bronchiectasis, bronchiolitis obliterans, alveolitis, sarcoidosis, pulmonary fibrosis, and collagen vascular disorders.

In the ease o;'cornpienterit“assoeiated hemolytic disorders such as PNH. CAD, and PCM, a medical practitioner veil i appreciate that C5 fragment Chit iby way of the terminal complement complex) contributes tagnifscamh' to the pathogenesis of these disorders See, c.g., Kaplan (2002) Gov Ophi Dnn;< M2.Ï 101/-2 3; 1 lilt (2005> i !ih kh- fienuh'f tAc·< / d; 1 I yd-dd-db and Router ei al. ι 200"! .\;<nn< /hi^c/mr/oyn-2:u 1 15. i 256-14x5; A, cord high, a medical praetiUonet mat elect m adnuntsiet one or more of the emi-dde antibodies described herein; m conjunction ith one ot name additional therapies lot the hemoivne dnordet such as a complement mhilntoi that ptev ents tbrmunon of the i '5h-° tennntal complement -„cmplcs ht ome embodiments til the methods described herein, the cmuplomenmo-somaKd disorder is not a complement-associated hetnoh. tie ib-msdes In some embodiments, an ;.eUo( '5a msribodv or .ui untigen--bmJmg fragment -hereof m adnumsiCiCd to a subject to treat, prevent, or ameliorate at lea-η i.uie sv mptoto of a a implement- issoriatod out utsukuorj -espouse ;e g„ the compierocnt-ussocialed intlammatms icsponse asp-.et of a eomplemem-asMa.ian.'d disorder* in a subject, hot example, m mui-tma nut mod;. deu rdvu herein can he it'-ed to tteat present, and fit ameliorate one oi nano symptom-- associated w pit a eomplement'-tssoeiated unlaromakvv. response snelt as graft rejection graft - v ersns-host disease ·.i i\ MD). lOperileoen injuries oog , to'dmv iug earchopulnmuur,' bypass ot a : issue transplant), and tissue damage following other forms of traumatic injury such as « burn (c.g.< a severe ben·;}, blunt trauma, spinel injury, or iiostbitc. See, c.g., Park at ai. i 1 *-><-)0} Λ mi l·*; 99(1 1.42--48, Tofukuji et a.!, i 1998},/ Thorne Caruiorusc Surg \ I 6t 6 s: iObO-,11)68; Schmid et ni, tI997} Shock K(?.y.) 1 9-17.4:, and Bless et at. (11><H> Am J Physiol hi some embodiments, on umi-C5a and body or an antigen-binding fragment thereof described heroin can be adnunistescri to a subject as a monotherapy. 4hername!>, as described above, the tout body or fragment thereof car; i;v administered to u subject as a combination therapy with another treatment. c g., another treatment tor a complement·associated disorder to" a caotpienioni-assocuued inikmirnatory :espouse, For example, die combination therapy can include administering to die subject ic,g . a human patient) one or more additional avers is ie.g., anti-coagulants, and-liy peri ensi\ox. or anti-mikmmatory drugs sc g.. -toroid-fi that pros idc a therapeutic benefit to a subject oho has. or i» at risk of des eloping, sepsis. In another example, the combination therapy can include administering to the subject one or more additional agents fe,g., as-: ano-tgh antibody, an a ms-11,-4 antibody, an anti-H..~5 anti body , or an ami-histamine} that provide therapeutic benefit to a subject who has, is at nsk of developing, or is suspected of having a complement-associated pm mum ary disorder such as COP Q or asthma bs some embodiments, an aun-i'da antibody and the one or more additional active agents are administered at the same time hr other embodiment-, the ami-CSa antibody is administered first in time and the one or more additional active agents are administered second m time, in some embodiments, the one or more additional active agents are admtrnstered first in time and the anib(.'5a antibody is administered second in urne.

An anti-Oa antibod;·, or an ami&amp;’ft-bmd*ng fragment thereof described herein can replace or augment a previously or currently administered therapy For example, upon treating w ith an ami-CSa antibody or antigen-binding fragment thereof administration of the one or more additional active agents can ccu.-e or dtminwh. e.o,. be administered at lower tevcis. In sonic embodiments, administration of the pre\ ions therapy can be maintained. In some embodiments, a prei tons the?any will be maintained until the level of the anti-Ce- antibody reaches a level sufficient to provide a therapeutic effect. The two therapies· can be administered in combination.

Mommrmg a subject (e.g., :¾ human patient? (or an improvement in a complement-associated disorder te g.. .sepsis. severe bum, RA. lupus nephritis, Goodpasture syndrome. or asthma), as defined herein., means evaluating the subject for a change in a disease parameter, e.g,, an Improvement in one or more symptoms of a gi ven disorder The symptoms of complement-associated disorders are we it know n in the art of medicine. in some embodiments. the evaluation is performed at least one C I) hoop e.g.. at least 2. 4, <\ S. i 2, 24, or4 a hours, in' at least I day, 2 slays, 4 days, 10 days, 13 days, 30 days or more, or ut least I week, 2 weeks, 4 weeks, 10 weeks. ; 3 weeks, 20 weeks or more, after an administration, the subject can be c-. -bunted in one or more of the followmg periods; prior to beginning of treatment.; during the treatment; or after one or more elemerns of the treatment base been administered. Evaluation can include evaluating the steed for further treatment, e g.. evaluating whether a dosage, frequency of administration, or duration of treatment should be altered, it can also include evaluating the need to add or drop a selected therapeutic modality, e.g,, adding or dropping any of the treatments for a cot η ρ i e m e a t-a v so c i at c d disorder described herein,

212ll .S3 fvAXèé:';: 2t2 . .ttf ft I., j ft tvlwftit Aef ΙΛ-.., Its .1 f.O T he disclosure also ieuturas thempeune and diagnostic kus containing, among other things, one or more of the anb-C5a a.mibodtes and or ontlgem binding imgmeuK thereof, described herent. The dmmpcuuc kits can contain, e.g,, a suitable means for delivery of the antibody or amsgen-hindlng fragment to a subject. In some embodiments, the means ts suitable tor subcutaneous delivery of the antibody or ammerHmnlmg fragment thereof to the subject, f he means cart be, e.g,. a syringe or an osmonc pump. That is, a therapeutic kit described herein can contain a .syringe pro-titled with an anis-('5a antibody or antigen-binding fragment thereof ie.g , n pen device containing the antibody or fragment) described herein or the kit can contain a pump (e.g.. an osmotic pumpt and one or more disposable cassettes configured tor use w itb the pump, she cassettes ρ re-on led with an awi-OSn amibody or antlgcn-brndutg fragment thereof described herent Oog., p re filled with an aqueous solution containing the aoti-Oa antibody or amigeu-hiudmg fragment thereof?. In .another example, the kit can contain a irans-sclcra! or tmphurtabie delivery device ic.g., a plug) that is pm-filled with (or otherwise eontams) a o.uuhcm containing a a a nti-C5a antibody or anhgon-binding itogmem thereof described herein.

In some embodiments. the means for deb venae an ami-Coa «m I body nr antigen-bindiug fragment thereof is a pen desnee ibrdrui; debvery, ht some. embodiments, the suesos is suitable for inimnnlmonary dedi\ery of dte antibody or antigen-binding fragment thereof to a -Object. e.y., for a an m treatment or prevention of a co m p; cm e n t - ;kso e i at ed pulmonary disorder each as, but not limited to, OdPD or asthma. Accordingly, the means can be, e.y., an oral or nasal inhaler i wo above). The inhaler can be, e.g., a metered dose minder (MD1). dry ponder inhaler {DPI), or a nehuHeer. Such a kit can also, optionally, include instructions for administering {e.y.,, self-administration of) the ami-f'5 a antibody or antigen-binding fragment thereof to a subject

The therapeutic kits can include, e.g., one or more additional active agents for treating or preventing a coinpiemerH-assoeiared disorder and/or ameliorating a. symptom hereof, for ex ample, the.rape.uik lots designed tor use tn treating or presenting a eompirmem-assoetated pulmonary disorder can include one or more additional active agents iin hiding. Urn not limited to, another antsbody therapeutic se.y.. an anti-igIf antibody, an unti-li.-d -mtihod) or an ami-lL-5 antibody}, a small molecule antr-lyf: inhibitor (e.g., montelukast sodium), a sympathomimetic fc.g , albtsteroi} an asnihtotic (e.g., tobramycin). a deoxyribonuclease fa.g , pubuozymci, an anticholinergic drug (e.y.. spratropitnn brontldei, a corticosteroid (e.g., dexamethiisone) a [badrcnoreccptor agonist a ieukoniene inhibitor te.g , :dlemon), a 5-hpoxy genuse inhibitor, a phosphodiesterase (PDf s inhibitor, a CDZ3 antagonist. an If.-1 3 antagonist. a cytokutc release inhibitor, a histamine HI receptor antagonist, an anti-histamine, an ami-inflammatory agent (e.y.., cromolyn sodium os' any other and-snflummatoty agent known in the art or described herein), or a histamine release inhibitor. hi some embodiments, the means can he me able for ismaoetdar administration of an anh-C5a anobody, or <tn antigen-binding fragment thereof, de sen bed bereut to a subject in need thereof e.y., a subject afflicted with AMD or any other complement-associated ocular disorder. The means can be, e.g,, a syringe, a trans-.sekrai patch, or even a contact lens containing the antibody or fragment. The means can, in some embodiments te an eye dropper. wherein the anii«€5a antibody er anftgen-btndtng f'hïgmeui thereof is fonnuUned Tor such arbnnstevuuon. buch therapeutic kils can ai so include, c.g , one or more add;donal ihenepcntie agents (or use m treating complement-a\w;e sated disorder of She eye. f he therupeude agents can te, c.g., be\ acwmnal· or die fab fragment oi bcaaeiaomab. rumbialmah. both sold by Roche Pharmaceuticals, Inc,, or pegaptimib sodium {Mut-ogeiwc; P(;;;er, Ine.) Such a kit can also, optionally. include in si rucooos no admens; ennu the ami-C 5a nan· body or anbgmnhhkting fragment thereof to a subject. in some embodiments, die means can he suitable for ;mmartscalar administration oi an am ini'5 a antibody, or antigen-binding Ifagmcm dr.neoi. described heroin to a subject in need thereof e.g., a subject· afflicted vciih RA. The moans can be., e.g,, a syringe or a double "barreled syringe. See. e.g , i :.$. Parent Non. O.0t6,f>45 and ;\0bH,6ii2. A doublo-barrcled syringe is useful for administering to a joint two different compositions with only one injection, iwo separate syringes may be incorporated for use su administer; nn dm therapeutic white drasi'ing off knee tluid for analysis trapping) m a push-pull fashion. Additional them pen or agents that can he administered with tine ami-C5ri antibodies or fragment1- in conjunction with the double-barreled syringe, or which can otherwise he generally Included os the therapeutic kits described herein, include, e g., NS.AIDs, corticosteroids. methomesate., hydroKyeltlonaqinnc, anti-TNF agents such as etancrecpt and iidllximate a B ceil depleting agent .-ueh a.s neuxmiaU. an intcrieukin-l antagonist, or a T ceil eostinudaiory blocking agent such as abataccni Such a kit can also, optionally include instructions lor administering the anh-C.'ëa antibody or antigenbinding fragment thereof to a subject. I? will be appreciated that, the disclosure embraces kits comprising one or mote of the artti-CSu antibodies described herein and one or more anti - intiamrouiory agents selected from the group consisting of N.VUOs, corticosteroids, methotrexate, hydrosycldoroqumc, anti-TNF agents -melt as etancrecpt and infliximab. a B cell depleting agent such as rituximah, an interleukin-1 antagonist, or a T cell costirnukttory blocking agent such as ahataecpi. The antibodies and agents can be, e g., JbrmuUtted separately or together. The kits urn be used to neat an inflammatory condition such as R A, Crohn's disease, inflammatory bowel disease, or any other infuurtmatory disorder known in the art or recited herein.

Also featured arc diagnostic kit*, containing the anti-CSa ami bodies or ammert-binding fragments thereof described herein. For example, the knx can contain a deteciably-labded form of an am f-C5a antibody {e g., an anti-CSa antibody or an antimouse CSa antibody) described ,herein for use in delecting or mtamitating the amount of CSa in a biological sample. in some embodiments, the kits can contain isolated CSa protein (cm., one or both of human and mouse if5a protein) and/or a control sample comparing one or both of human and mouse C5a protein, in some embodiments, the kit corn urns a multi-well plate coated with a first arm-CSa antibody having a first spectficirv. The kit also commas a second anti-CSa antibody (e.g.. a dctectabiy-labeled second anti-CSa antibody) having a second specificity. Such a kit ts designed foe use in caplnring, with the first antibody bound to the plate. C5a protein (e.g;., human CSa protein) in a sample fe.g., a biological sample) contacted to the plate and then detecting the captured C5a protein using the second antibody. hr some embodiments, diagnostic kits include both an ami-mouse- CSa antibody and art anti-human CSa antibody described herein. In some embodiments, the diagnostic kits include an anti-CSa antibody that binds to both mouse Co a and human CSa, i’he MIóWMg examples arecibiendy: ίοηΙΙιικίορο, Βοι !i:mitvfh.e mvebfiom·

EycutpSes i he presently described ami-CSa antibodies are heormmed forms of marine antibodies generated under the to Ho whig immunization protocol, Immunizations to raise antibodies against hinnan desargmated CSa v>ere performed on font' mice including two mice of the -main DBAabj and two mice of flic strain Λ .1. These strains woe selected because they carry the allele Hc'\ which makes them deficient In endogenous Cm Alt immunizations mere repeated at 14 day intervals tor a total of three immunizations. Ail enlmaK received a subcutaneous booster mmumm.mon of approximately 50 ug of purified CSa in TOO μ l of adjuvant emulsion approximately U| days anerthe last u'nmntmmmon and 5 to 7 days before harvesting. Titering of scrum bom immuni?.ed mice, using an I'lLISA assay, showed chat the mice exhibited a strong antibody response agaia^tlMe Οίκηση desaqpnated CSaimmmiegeRv. lAiuncteJLJSsicffiiü^ Λ subset oi tree mouse anti-human C5a Fobs that w ore rcprcsmmuo o of neoepltopv seBeio c hubs were com ened to fuU length mouse MkBa antibodies designated as funO-MMl:, 5sn \ 01 Mb', Sant 'ΜΜη, 5an; 79Mb. and ban skOMt. I hese antibodies were evaluated to* y-peeifieoy using Bioko,es interferometry m; an Octet {HorteBso iitck! i i he amino acid sequences of the light chain and heavy chatn CPU sets oi each antibody , as defined sty Kaibal. are set forth ei Table 3 ; Brie sly. hamen C5a. human 05a des -\rg, human full-length 05. or 05a pa5’a logs human 03a ansi lutmass 0-7¾ were conjugated lo biotin ai a stoiehiosnetry oi' a 1 (biotin}. 1 (antibody} through amine groups -md i mmole lived on a sireptaudin tsp. i.oaded lips a etc. tliett exposed u> a solution containing 30 nM of umi-Oëa IeO antibody. hack of the antibodies bound to C5u and desargmawd 05a Mane t>f the «nlt-l 5a 1 ei» unbbokus botntd lo C5a or to ( -la. However, 5;.tn I 7 k Mb and 5;m I 7C!Mfc ouch bound to full-length hem an 05. A -wnsil amount of binding, was observed between .MnOdsMb and ! hi Monet h human 05. I hm ever,, the binding of danO-lKMir to 05 was much tes-> than die binding observed to 05a.

These resulis confirmed that mouse anti-human C5a antibodies - 5an04fcME, 5ani0lMI:.„ and Ptmt^OME - hound lo a neoepilope on 05.¾ that was occluded in native, full deugd; (55 os' generated after the cleavage of 05 into fragments 05a and (55b. 1'he results also indicated that the three antibodies were selective for human 05 a as compared k.> para logs t 3a ur 0Ό.

Xii.bO.3.... Aptmo g\cyd .Sequences..fory fuvc MluMnc .Anf<75a

'81\ ro KoL ïolois ^ H) \u' {.'PR umtno acid sequence defined according fo Rabat ei al i\n>tr<i). Λ serie* of sandwich assays were pedonned by Octet on the selected subset· of mouse anti-human 05 a fgCOa antibodies to determine the degree of overtap of the C$a epitopes for each of the five representative lg.G2a antibodies. Briefly, a first antibody was biotinylated and immobilized on a streptavidin coated tip on the Octet platform.

Next, human €.$&amp; was captured from a .:.0 nM solution on the immobilized antibody. Tire tip carrying the antibody-C5a complex was then exposed to a solution containing 20 nM of an unlabclcd second ami-CSa igG antibody. The elicitation of an additional association profile in the scnsogrsrn would indicate drat the two antibodies bound C5a simultaneously in a ternary complex and that the binding epitopes tor the two antibodies were non-overlapping. A failure to obtain a second association profile upon addition of the second antibody would indicate that the two antibodies bound C5a in a competitive mannen l.e , the epitope on 05a ro which the second ami body bound was occluded after binding by the first antibody, in contrast to rxm--competitive binding, competitive binding does not necessarily indicate that the first and second antibodies recognized the same, or even overlapping, epitopes on human C5a. Using tins approach the binding sites of the five representative anti-C5a antibodies were assigned Jo <1 distinct epitopes on human CdalFig. Π Antibodies dfiniMKME, San I HOME, and fan Ai 1 Mb competed with each other. While danOds and San 1HO also compered with the non-ncoepitope selective Senl MMF:. fan 101ΜE did not. which indicated that SanO-IKME and 5an I &amp;OME recognize a ooocprtope that ts different than the epitope recognized by SanlOtME In addition, while the non~neoepitope select'd e antibody fan 1 MMe compered wish non-acoepitope selective Sani7b1 ME, only the latter competed with San 1SÖMR and San04^MSA showing that 5ani "9ME. and hen i 78 ME. bind different epitopes that are accessible both in C:3 and f’Su. The results also hidieaie that scene combinations of da; antibodies could be used in sandwich-based assays to detect and-or quantity the amount of (. ha in a sample f.M.VIi i.l .Si..:...: y. ill .bTEi.i /. ΪΛ>.ΙΛ. .M.f'. 1 S'.tV.f.. bl.\f(t:\ ΓίΜ.h t j.] tyan.. C'. A . .< I n.1ΛI.': Q.sl.t. h'.ir

Ine variable regions of two related motive ami-human i.'5a antibodies -5an KM ME and 5ani 85ME - were selected for Stumaoizarion as full length iuO antibodies. I himantzahon oflighi and heavy chain variable regions was based on identifying individual framework regions from human antibodies {with a preference given to germ line v-genes) with a high degree of sequence identity to the original omring parent an a hub >, Methods for Mcmifvwy sitituble candidate framework regions are described in ITS. patent no ?J95.*'4X to Rothcr and Wu. Definitions of framework (TWt and eomplcrnentarity determining regions {C'DRs} were performed according to methods described by Kabat, Choi his and I MOTam International Inimu-toGenciics Information System. France). Briefly, database nueries were performed independently for both the light and heav y chain variable regions wiih a variety of antibody fragments including.- intact murine variable region from FWI through PW4, intact rnururc variable regions excluding ( DRs and ai! possible fragments of murine variable regions including one. ολο or Hire.':.· Damcwoflm wuh or w-thou? their flanking CDR x, Human frameworks were selected from this Candida ie gooi based on their overall segue nee identity to ihe original murine antibodies and fioginenis ihcicoh Routine molecular biological methods were employed to assemble small combinatorial libraries, of less ika» 1tV members. in which each set of murine CDRs mere flanked by ail possible combinations of seise led human frameworks. Ihese humanised antibodies o ere es pressed as soluble fads and evaluated lor buiding au ocsiugoiaied C:5a usmg ELISA, f abs iteii bound to brui wem then subjected nu DBA sequence analysis. from these binders a subset of sis human seed Fobs were reformatted us full length I gibs ihuman IgGc or human lgi.12 Sbib Additional munani/ebon was performed in two antibodies i BNj.Vt 1 and BN jib! i b\ repiacmg murine residues In ( DR 2 of the light chant wnh their eonesponding human germ bn a. amino acid' The amino acid sequences of the Immunised anti-Oa antibodies BNlabd. BNU36?. BN )57 I. B\U3?S„ BN 33 bo, RNJooo, BN.big i, and BNi.lsg · are set tbah in "fable 2 above.

Aobfilllb.d.....12etcrnnnjn^ghe.aΠ\]uty·_ oflütf bumanjgca antBhnntmyC5;ga>niKuh 05 u

The humanized antibodies were subjected to BIAeore analysis to quantity their rcspcctix'e affinities for human C5a. bee, cop. Karlsson and Larsxon (2003 t Methods Mo! Bio· 248:38^-415 Briefly, each oi the humanized antibodies were screened with 5-4 concentrations of human C5a iantjgcn) «sing a capture technique. The antibodies were captured by an Ami-Ec (human) directly immobilized on a CMS sensor chip with v arious concentrations in the range fforn 0.6 nM to 5.0 nM of human OSa passed over the -ensor chip surface. The surface was regenerated w ith 20 m.M HOC 0,0Tb· F20 after each cycle to rctnov.s bound antibods and autigett. The data was e\ Turned using Biacorc BIAev ehtniton so go ore using a 1:1 Lanemmr Mode! Fit (Umax. Globa! Fit: PJiLoca! Fit); Kinetics information sudt as (kg Association Rare constant), iky Dissociation Rate constant t and Ks i laquiburiuso Dissociation constant) was obtained from ihe fib The results of btc analyses are set forth tn fable -!. These experiments were for screening purposes with a minimal number of analyst, concentrations {3 to 4 ί with 1 duplicate fcbe approximate Moetics Fable 4. I^g.l:„jAtMaJdcg^^

All of the fmrnanieed antibodies specifically bound to hunt an {'Sa with a K:< less than i .20 nanomolar. All of the antibodies with the exception of BN)371 bound to human C5a with a Ko levs if; a η I nanomolec. Three of she amihodies. B'Nj AVf BNjph I, and BNjab.3 hound to human €5n with a Ke leas than 100 pieomolar. .fiMOifiki..... Λη m vitro oeuuoplnl activation assay was used to evaluate the activity of the humanized antibodies. I he assay is generally described In. e,g., PacttkowsU et ah t P>0;>} Bf J PiuinniH.oi l.wh(7j;!4o! -1400, end serves to qua nutate the amount of myeloperoxidase (MPO) produced by neutrophils as.a measure of neutropfdl activation Briefly, poly morphon nc! eat ceils., the majority of which being neutrophils, were isolated using density centrifugation (mono-pols resolving medium catalogue number; 9169X049; TIP Biochemicals; Solon. Ohio) from whole blood trom a healthy donor. The celK were '.wished once wtth phospbaie-bvdTered saline iPBSt and she red blood ceils tRBi. ) removed from the cel) population by lysis m a hypotonic solution (AC K lysis buffer catalogue number 10-54X1); Looeas. A tier two more washes wtth PBS. the RBC 'free celts were resuspended et a content; at ion ei'4 s; i 0' eclis'tnL in Hanlt's Balanced Salt Solution (HBSS; Mediaweb, catalogue number: 3 1 -023-CV?, which was supplemented with calcium and magnesium and further supplemented with 0. PM gelatin (Sigma Aldrich; St, Louts. Missouri ï {heretrutltcr the assay bullen.

Cytochalastn B ; Sigma Aldrich) was added to the eed suspension m an amount sufficient to reach a concentration of SO go, mL. lire suspension was then incubated for 10 minutes en 37"(€ 100 oL of cede was added to -acid' of IMxttfomed ;!i'wei 1 plates. The wadis of the plate were grouped into several different sets, fa eh of several of the different, sets of webs contained an ami-Clin. antibody each well ol net I contained a humanized antibody that binds to une leaved, native C5, but not to free €:3a; each well of set 2 contained the BN ,1367 humanized and (€;;. and body: each wed cd' set 3 contained the BNJ369 hurnam/cd and-CSa antibody; each well of set 4 contained the BNJ371 humanized unti-i'eu antibody; each well to'set 5 eon finned the BNiMX humanized imfi-C5« antibody: each v\di of set 6 contained the BbJ.181 humanized anft-C da antibody; and each well of set ? contained the RNJohd humanized a»h-{.'3a antibody, f ach welt of an eighth set of weds contanten no antibody. A sartge of antibody concentrations was evaluated in each set of welts, the range including 0.0* nM, 0.4 nM, 2 nM. and 10 rtM amihody, C;5a (obtained front Otmtplement Technologies, Inc.) was evaluated at a concent ration of 3 nM. Λ 1 OX working concentration of 20 nM was prepared in the afot'ernenhOHcd assay buffer and 20 μΐ. was added to each web. After addition of Of a to the wells, the plate was incubated for 10 minutes at 37 '€. following the incubation. *"Ό ub of PBS was added to each well of the plate, ihe plates were subjected to centrifugation at 1200 rpnt (approx;amide 3 35 x gj for 10 mmutes at room temperature. 100 μ 1.. of the supernatant from each well was transferred to the corresponding well of a second plate. 2:5 pi., of substrate I Sigma Aldrich catalogue number 1()44() t a as added to cad; well of the second plate and the peroxidase -unction was allowed to develop for approxImmely two to five minutes, ihe reaction was terminated by ihe addition of 25 el of 1N I IC I. The OD at 450 nM was recorded.

As show a in Fig. 3, ail of the humanized ami A pa antibodies inhibited nctsirophi! activation in €mw These results indicate that the Immunized anu-Coa antibodies described herein arc potent inhibitors of C5a-rned$uted .signalseg in vitro and support the conclusion by the irneruors that the aoubodics are useful for Trcaiinga variety o! complement-associated disorders (e.g., complement-associated inflammatory disorders) in humans. lidltVIH'is. iLOhargctengatw^ o f a sti Λ series of sandwich assays wore performed on a selected mouse ami-mouse C5a IgG antibody 5 an 195MB · to determine the specificity of the antibody for (i 5a.

Briefly, the wells of an assay plate were coated with the fan i^SME antibody, the plate was washed thoroughly to remove unbound antibody. Nest, wells' containing fan i be ME άere contacted with mouse Cda for a time and under conditions sufficient to allow the antigen to bind to the antibody. Unbound protein was removed w ith washing. Follow ing the wash step, the wells were further contacted with a solution containing a second, hiotiuykted ami-Cxa antibody. The wells were again w ashed to remove any unbound second antibody. The amount of binding of the second antibody to diet well was cpumbiled using strepen idin-ionjngated horseradish peroxidase t HR If) Hi·;· amount of binding of the second antibody was ; function of the binding off da to 5 an 1 95 MB.

In a parallel experiment, a set of fee195Mh-eoated welts were incubated wuh ft.ilMength mouse C5 protein, rather than 05s. Follow ing a wash step, the wells were contacted with a solution containing a second antibody: a biotinylated ams-mouse 05 antibody Fbe amount of binding of the second antibody. ss 3 function of the amount of 05 bound by Pan 1 <;OML was quantified using the strepsavidin-conjogated HRP too-cruet. A lack of binding of the second antibody mdiea;cs that dan i 95MB does not bind to cud-length mouse 0:3

While dun 195ME hound to Cds in a dose-dependent manner. no binding between the antibody and fulldetigth mouw (.'5 was detected using this assay These results indicate dint 5an I95ME bends to a neo-epitope prescig in (.'5a

The tektivo binding affinity of 5rm 195 Ml Π for mouse C5a was fun Iter quantified using Bl Acorc. The kinetics oi' Pan 19501F w ere measured using a capture technique.

The antibody was captured by an Anu-Fc (mouse) due·, ih irnrnobibvted on a CM5 sensor chip wish rariotts coucetrt'cattotis between and inclusive or'0.4 nM and 25 nM of mouse C5a passed os or the sensor chip surface. Dupheutes ot each eoneetitration were also run.. The surface of Hie chip wus regenerated with 10 mM glycine· HCI pH I ? after each cycle to remove bound antibody and antigen. The data were evaluated using B-acore Bl A ova I nation software using a HI langmuir Model Fit iRmuxtOlohal Fit; Ri.Local FiiT Kinetics information such as k., (Association Rate constantk.; (Dissociatum Rate constant'! and Kp {Lkphlmnum Dissociation constant? was obmisied Bon· the fit. t he results of the kinetics analyses are shown m Table 5.

These results indicate that the mouse unit-mouse ('he antibody is not only snt-etftc for C5a, as con reared k> iuti-iengih mostse 05., but a iso that the antibody binds with high affinso.· to mouse 05a

The 5aol95ME anti-mouse i05a antibody was evaluated m a mouse model of cuilagen-mduccd arthritis. Male DBA I f.aej mice (0 to O'? weeks old) were immunised by iutraderma! injection at the base of the tail with 300 ugoi bovine type Π eoUagee emulsified w ith equal volumes of Freund's complete adjuvant ihc procedure was repeated two weeks alter the first immunization. Mice were inspected dads to identify inflammation re an initial knee joint. Once the mttiet mSlemnurkort was idem died, mice were inrraperitouually adroinistcuod three' ume> week the Part S05ML' auti-mouse {.'5a antibody (40irw. kg? or a control antibody {;lO;ngkgk The thickness of the initially tniimned joint tin rooft was mensured daily to day 12

As showt? in Fig. Ik SartHOMF. reduecd knee joint thiel-.ness as compared to Utc control antibody, 5an 195 ME appeared to provide the benefit, of maintaining a knee joint thickness below 4.5 mm.

In addition to evaluating the ability of the 5anl9>ML antibody to redt tee swelling of the initially inflamed knee joum. the ability of the San ; 95M E anti-CSa antibody to prevent migration of inlntntmaUon to new joints vs as also evaluated. The number of newly recruited .joints was meant red daily from day \ to day j 5. The results of the experiment are set forth in Fable o

Fable 6.....EOlhAtcyof 5uni95ME^

As shown h\ Fable t\ rntee neared w-dt 5»nl95ME hud markedly tower newly inflamed joints as compared with control Ah treated animals by day 1 2. San 195Mb· treated mice also bad on average markedly fewer newly inflamed, joints

The a (threw in the mice was also monitored and defined using a clinical scoremribriox index, bach lints? was graded daily according to an established scoring system (Cf normal jotnt; L mild/moderare v isible erythema and swelling; 2* severe erythema and .swelling effecting an entire paw or joint; 3, deformed n;m or joint oith ankylosis.). with a maxim urn score of twenty-four per animal. See. c.g , Vv'ang. et at. {20001 ./ /newnam1 161:5340-45-47 As shown in Fig 4. mice treated w nh the ano-nnmse CSa tmttbody 5an fAb-IF exhibited a marked reduction n; clinical score taxes age score of leas than 1),. a-, cornpared to mice treated with the control antibody {av ersge .score ;mov c 6 k (>ver the course of the study.

In sutmnary. these results indicate that the surrogate anti-mouse C5.n ts eflecttse in tnndtng R \ both at an utittal jomt and the sow moon of inflammation to secondary joints tn the mouse model of disease. The results also strongly suggest that a therapeutic ami-human f'5a antibody, such as any of the humanized ami-CSa an a bodies described herein, ia useful for treating humans with RA. A human patient is identified by a medical practitioner ns having rheumatoid arthritis in a single articulated joint. The patten; tu shortly thereafter admen sirred immanividany or inf mperi tone»! jy a composition containing a human» trod anti»C5a antibody described hem as in an amount sufficient to reduce CSa-vnediaScai (.'5aR t signaling locally v, stitin tin: somt apace. 1 he patient and medical practitioner observe a substantial unnrovetueni in at least two known symptoms of rheumatoid arthritis following the treatment. The pattern receives intravenously administered "maintenance Joses" of the antibody every month to prevent reoccurrence of the symptoms, to prevent, the progression of RA at the single joint, or to prevent the migration of RA .symptoms to a second joint. hxaiywl.e 9, Use oftm untl-(;ên amihodv to ireatsegsjs Λ human pattern ο identified by a medical practitioner as having sepsis. The patient is shortly thereafter administered a composition containing a humanized ami-CSa antibody described herein at a dose of approximately 000 to 000 mg by wav of intravenous infusion. The patient and medical practitioner observe a substantial improvement in at least two known symptoms of sepsis during the treatment. The patient receives intravenously administered "maintenance doses5' of the antibody every two weeks until the patient leaves the hospital. Λ /.t i 1.41 .Vs.T.tlct AtVt Aê:. t.V.tt Λ human patient is identified by a medical practitioner us having a se\ ere form of COPD Once every two weeks for four weeks the patient is administered a composition containing a humanr/.cd ami-CSa antibody at a dose of approximately f>00 mg to 900 mg by intravenous infusion The patient and medical practitioner observ e a substantial improvement in at least two known symptoms of COPD during the mica! treatment. For example, the pauenf receiving the sml-CYx sub body has a reduced frequency and or severny of COPD-rclatcd exacerbation.*. The patient continues to receive mtra venousdy' administered 'Tnaimcftancc doses" of dux antibody every two weeks to mam mm the reduced frequency an&amp;'or severity of COPD-rclaieJ exacerbations. A b:.tman patient is identified by a medical practitioner as having a severe form of asthma. The pattern ts prescribed a therapeutics humanized antt-C5a antibody io be administered by way of an inhaler. During the next attack of brontbospasms. the patient seif-ad monsters the anil-05a antibody in art amount sutYicicrU to reduce the € :3a-roediated inilummatory response sn the lungs of the patient. The patient continues to use. the inhaler as needed to prevent or lessen the severity os'asthma attacks

ItddmnkjJ......AMüfofoLdfoxkYddfofo

Sm era! additional xmuxsmm mere obtained front the communed mice <*ee. ifxarople I) attd hart her identified by if I. ISA as capable of bindurn m human 05 a. The additional antibodic.' include 15 unique light chair: CDR sets (set forth tn Tabic ?> and 14 unique heavy chain (T)R set-mas set forth in Tabic fo U mqu^ L .an dK abamde fit bfoi^iOlfon

'"SIN"' in the Dsble refers io '"SHQ ID NO " * CD I? arvnoo scul seeueüucs ere defined recording ?e Kulun ei O. i \Np;\n

Table S.....Amino ..Acid .Sequences of &amp;yerafUnin^^ CPK scqnences fx>n: /\dduli>n^d Slurine An?hhu;ADcC:0 Amibrniic:;:

"SIN" in me Tabic reicr.s to "STQ iΠ NO." * ( DR aouno acid senacnct;* are (kilned .according 10 Kabm el ai. V j and v u puirmgs giving π so to the 5a.nΠ/Vff·.. 5an054ME. 5an: Ki'vffi, :knil KSMF., éanl^lNik. ami 5emK)7Mi; uni-bodies are evident bom Tsides ^ and 8. Additional exemplary pairings of the heavy Drain and light chain van .Ole regions' and or CDR seis me set fonh in Tabic b below I'miieyfAnumyAQdikguencesJ'or/Xdmuona^fdoiSM^ 0>E.Sco

"SIN"' in the Table refers to "HEQ ID NOS' * CDR amino actd moucnce^ arc defined according to Rabat et a), {supra).

Es<lSH>is,L^:,„S;jdS:S>lSlLh*:S]]SSliS'R.‘SMi:iSlSRO.i-^y„iSSA^Sii.'SS'

Several additional humanized &amp;r>ti-C'5a antibody heavy chain variable regions were generated, ell of w hich when paired with a common light chain variable region (the Itghi chain having the amino acid sequence depicted in SEQ ID NO. 16) bound to human CSa with a ICo of less than 1 t»M as determined by Blacore analysis (see above for methodology r All ofthese additional humanized antibodies bound specifically to human C6a. bin did nor bind to native, fully-folded human C5. (Ala. or CSa as determined by Octet analysis (see above for methodology } The additional humanized antibodies corn:-· mod; fnheavy chant variable region framework teg I on I commsnsm one ot me Iblkm tug arnino acid sequences; QVQf VQSGAEVKICPGASVfCVSCK. ASGYTI-'T iSCQ ID NO.OHi or QVQiAQSGSELKKPGASVK VSCKASÓb'TFT (SEQ ID NO.iGn (sit a heavy chum variable region framework region a containing one of the to Ho ml rm amino acid sequences: WVRQAPGQGLEWNtG (SBQ ID NO:?0? or \VYRQ ASGK.GI.F: WVO tSiëQ ΙΟ ΝΟ:?ί): i Üi> a heuvy chain variableregionfemgwöffc region 3 containing one of tbc Hollowing asni.no add sequences: RVTtTRDTSASTAYM(DASLRSPDTAVΥΎΓΛR (SRQ ID NO.72K R ΥΊΊΤ.ΛDfSTSTA Y Mf.LNSL R.Sf:.DTA Yb'YCAR (SEQ ID NO:?.j); Of R YTiTRDRSMST A YML'fSSLRSPDT.AMY YO AR (SbQ !Ü NO.74K and a heavy chain variable regio;·? framework region 4 eenkennig the following ammo acid sequence. YYGQGTfYTVSS (YlïQ ID Ni 0:75), Exemplary add nier; ai humanized heavy chain variable regions comprising one or more of the uddhional bnmanmed traineeork sms described is; this sec eon are set forth in Ion. 5. ingnmO j,3.....i Ac of inianOrhhOMliCha amib^ A murine modal of t'5a-ncinropcnia vs ns utilized to evaluate the efficacy of an anti-human free (.0,: antibody /;; wYo, To induce neutropenia, purified, native humsn C'jü (h€5u? svas administered by way of intravenous tail vein injection to Baib 'c mice. The number of circulating neutrophils was evaluated up ;o fo e minuses after admtnistomon ol’ltC:5a.

Administration of 300 pg'kg of hC5a was consistently found t.o induce neutrophil activation as measured by the myeloperoxidase «MPOt release assay (see Example 5 for use with serum as compared to cell cuds;re supernatant, uipnnsnd neutropenia fa red net ion in the number of circulating neutrophils), tn addition- plasma levels of hC5a and the human ami-CSu antibody BN.I383 (when administered to the mice, m/ru; vvere also measured to establish the pharnmeodynamfc response (see below).

Peripheral blood neutrophil counts were examined before challenge with hOSsor vehicle control. Compared to sham-treated control mice (1.37 ± 0.00 x ΗΓ rnl.. ?, neutrophil counts in mice treated with anti-human free C5a antibody (),32 e 0.13 xIQ1' per mi. at 24 rng.-kg; P :=- 0.05 t m isotype control rn.Ab: l .31 ± 0.10 >: ΗΓ per mi. at 24 tng. kg; P>0.05> remained the same. These results indicated that amibody alone did not induce changes 1st circulating neutrophil counts.

To evaluate the efficacy of an anm-CGu antibody to inhibit hC 5a-induced neutropenia in mice, different dosages (24 mg/kg. 12 mg kg. 6 mg/fg, arid 3 ntg kg'? of the anti-human C5a antibody BNj383 were administered to Btdb c msec 24 hours prior to hCSa injection. Administration oi the arm-€5a antibody 24 hours ahead oi hC5a allowed iorthe pharmacodynamics properties of the antibody nr be studied during ihe p-phase of antibod) elimination from Hie mice

As shown m Fig. 6, neutrophil counts after treatment are expressed as a percentage of “baseline" (where the count at time 0 equals 100%). In sharn -treated control mice, the neutrophil counts -were 7s).02 .4· 5.71%. 67.42 4 3.232-,. and 39.54 ± 2,: I % of baseline at i, 3 ansi 5 minutes post h€5a administration, respectively. The Isotype control antibody-treated mice denaonsnauen a significant reduction (Pu).Ol} in the neutrophil count to 6.76 ± 0.8!% at i minute. 6.6b ± (),81% at 3 minutes, and 8.29 ±. 0.79% at 5 minutes alter imiavcnous injection of hC5a. The ants-C.aa antibody exhibited, a dose-dependent effect on hCen-indueed neutropenia. At the highest dose, 24 mg/kg, the ami-CSa antibody completely blocked the neutropenia. Neutrophil counts were 70 35 ±. 8.64% at I minute, 63-85 .:. 6.085¾ at 5 minutes, and 59.65 * 6.5 i% at 5 minutes, vs inch, sea* comparable to the neutrophil levels in shurnureated control animats at the same time points. Fhe lower doses of 12 mg4g or 6 rog/kg of the anti-OSn antibody also si gin Pcam iy inhibited neutrophil depletion (12 mg/kg: 42.61 ±. 5.1251, at I minute, 45.88 i· 8.29'··',. at 3 minutes, and 41.02 4 7.080¾ at 5 minutes, P- 0.01. 6 mg/kg: 18 00 4 8.8 at 1 minute, 20.20 ± A.44% at 8 minutes, and 28.03 4 4.51% at 5 minutes. P- 0.05} followmg administration ofhC5a, as compared to the isotype control antibody group ib.7o .· 0.81% at I minute. 6.68 t 0.81% at 3 minutes, and 8.29 .?.- 0.7953 at 5 m none .--4 The ssotype control antibody is an antibody that binds anthrax protective antigen 63 and contants a human lgG2/4 isotype Fe region. Administration of the lowest dose of the antt-€5a antibody (3 mg. kg; did not significantly reduce neutropenia (6.28 r 0.88% at I minute, 6.71 4. 2. 14% at 3 minutes, and 8.75 o 28)80¾ at. 5 minutes, P-0.05r Set Ftg. 6.

Anti-human €2¼ anti body Inhibits hC5a~indumt VIΡΟ release in vim

As discussed above, human C5a ucto-ues neutrophils through eross-reaettse binding of fhe mouse C5a receptor. Myeloperoxidase {MPO) release is a consequence of neutrophil activation through C5a Finding to C5ak See Darren ct ai, {2004) Mo! Piuofn [5^1):869--8/9, intravenous injection ol;ret.ornbinant human C5a Into the mouse can induce neuoopenia arid activate 5icutronlii 1« in cireukitiotv. The ability or ar; amt-human free i’’5a antibody sc inhibit MPO rdea.se clue to neutrophil activation <n v;Vo w as evaktated using rise anti-OSa antibody to hind free hi" 5 a and prevent binding to the murine C5aR.

An in r.:v.·,· experiment < in which hitman C5a was administered to msec) wax pertormed as de sect bed ;Tuvc. The level of plasma MPO at time 0 w as 50.25 v 22 ng’mL in the sham-treated comas! gamp, hoc mnunvr after intravenous mseuion of x abide bn Oer., MPü koel·- wem not sigmftrsosk ΟηηοηΜ (“7.4b e 2; 21 tot ml.. P 0.05b Prior to mtrav enous mscetson of hi.'5a. MPO ko els in woe. pc cosnrol andbody -treated ;75.;7,·λ· M bo ng-su1..) or anti-Cva antibody-treated animals (bee" ·λ· i -i.Xo ng/nsl 5 ‘Λ-eve comparable o ah the levels observed it; the sham--winner: control auimtils After intravenous injeetion of hf 5a MPO leads at ait doses of if 5a wen: raised and remained at signsficantly hteder leads at 5 minutes (leg, ?).

When eompmvd to issnyna conti id amibod) -reared animals <221.00 v 51.02 tee-ft! C the ;wh~hC5;s antibody-wealed animals shoveed dose-depetidea? reduelkstt of MPO levels < 11 -1.23 -t 25.2b nadab. P- 0.05, in 2-1 mg kg cohort: 10--1 00 a 25).03 ne mi, P<0.05. in 12 rug kg cohort; and 126.00 a: 3b.40 ng.dni.., P;:::0.0X, in b rng.’kg cohort) The MPO levels oftovv close O rngdkg) a«ti-C5a antibody-meatcd animals i 17b 55 ±- 23.05 ng/mL I were -tot signlfteamly difiere-tt from isotype control antibody-treated animals (10-0,05),

An an)i425a antibody reduces circulating M. Sa levels 1st os lee

As noted above. (25a is « potem snilammatory peptide with several biological junctions. Thti.se above studies devnoststmted that human C5» cros.vreacis with murine CbaR >.>n neutrophils since intravenous injection of recombinant human {'5a can induce neutropenia. While thts dtseOsurc is in no way brnhed by arty particular theory o? mechanism of action, the a ns 5-(25 a antibody may be inhibiting human CAwmdueed neutropenia Ivy forming complexes with nf.'fa and preventing hC 5a from binding to the muntte C5aR expressed on the cell stuf ace, hC5a levels were measured its the plasma oi mice before and 1, 5, and .5 minutes after intravenous administration ofbCSu to confirm that the effects of the ami-h€5a a au body in vh'<> were doe to ns hiodioy-tkpendcnt hihibhiost of h€$a.

An experiment was performed as described above ustng the monso-hOSa-induccd rwuiropeoia model. h05a was nor detected m plasma io am group of mice prior to administration of hf'5a at time 0, using an enzyme-baked immunosorbent assay {LUS A) The ievd ofhCSa in plasma, however, increased to a peak at 1 mm me * 7783.50 ± 327.7¾ ng'mU. then reduced to -478838 * 260.0 i ng'ml at 3 minutes, and then to 3855.7$ * 2V8.*/> ng-nu. at 5 mmuru» tot towing inoavertons tnjocuon ofb05» (tn isenype control mAb-treated mouse). Compared to control ami body -wonted mice, the level of hCda in mice treated with 24 mm kg of the awi-CSa antibody exhibited a 43., 30, and 2 Vfoid decline af I numrte (178.4 i 14.)4 ng/mL), ) nurvutes {15s.4 .-.. I0.43ng ml κ and 5 minutes Π<'<7 2 -t .15.6.1 ng mi.), respectively. The level of hC5a in plasma worn the 12 mg'kg and 6 mg-fkg cohorts were 235.0)) a. 22.3) and 600.20 a. "8.73 ng-ntL at 1 minute. 2 i0.80 .t 19.$» and 527.o0 £ 53.25 ng mi. at 3 mintucs, 102.201 7.40 and 50$,00 £ 45 90 tig-mi.. at 5 minutes, respectively. The anft-C5a-treated mice exhibited signineamly reduced hC5a k-vcls in a dose-dependen· marmer during neutropenia following intiavenoas inject sort of hC5a {P -s 0.001) Although dte mice tecois tug the lowest dose of ami-C5a antibody t3nsg.'kg) were not spared from hC5a--indueed neutropenia, the mice nonetheless had a significant reduction It? plasma hf $a 13130.40 i-433.5-4 ng/mL at ! minute; 1932.00 - 268.92 ng'ml at 3 minutes; i593.00 ± I09.6k eg mi. at 5 minutes) as compared to plasma hr5a levels ibmtd in iso type control antibody-treated mice {P --0.05). ,Vt- f ig. 8. These data indicated that administration of anti"0$a antibody to mice significantly decreased the conconuation of wee hf'Sa in plasma. resulting ttt greatly ameliorates! hOSa-indaced neutropenia.

Taken together, these results presented in this section twite a ted that an ami-human C5a ami body described heroin can inhibit the biological effect ofhuman C.'5a in an ir; vao disease setting and provide strong evidence that the antibodies {and antigen-binding i ragmans thereoOare useful itt, among other things, treating or proven ling complement-asoac sated disorders such as any of those recited herein 1 : j. ί:".. 3. ."I.:... .Δ .1 LrJ'A! :5. .1?AilVi.. : ..ί·.ΐ1. J'A: ΛΪ.5 Λ Is .^Ιχ . .V. >Λ .3.ΐjj.. λ :. .S λϊ .. ;(ί.ί;ΙΐΛ. .Vi.^.TAr. L\ .Vi λ!\ i'A ;Λ. Χ> ΟΙλΙ .^.

Several humanized ami-bCSa antibodies were tested for thoiv ability to crossroad with C 5a from one or more non-lnursan mammalian species. As noted above., die benefits of sued am suii-Cfa antibody are numaroua, e.g., the anility of a research or medical practitioner to model the efficacy of a therapeutic ainf-CSa and body in a non-lusmust disease model prior to administering the antibody to humans. Testing in non-human mammals can also allow for determination or approximation of the appropriate dosage of an ;ujti~C5i·· antibody required for efficacy m humans

Briefly, BNBrAL 8N.B66, BNjfó-T and BN.13S3 tdescribed above) were evaluated to determine whether they could cods nmunopreci pirate C5a protein in the activated serum from several non-human primates including baboon, rhesus macaque, and cynoroolgus macaque The serum was activated by addition of zymosan. Following, an overnight incubation of each antibody with activated serum, the antibodies were separated front the solution phase using protein A-conjugated agarose beads The beads were washed thoroughly and then boiled in sodium dodccyj aid tie c - po I y ae r y I rum J c gel electrophoresis (8DvPAGE> sample buffer containing jf-mereaptoeibanol. The boded samples were then subjected to SOS-PAGE, Non-human primate C5a was detected by western blot using ihe commercial iy-avaitable anti-CSa neoepuope antibody P2P4.2 (Abeam, Cambridge, MA}. Each of the antibodies tested was capable of tmmunoprecipitali ng f5a {or CSa desarg) from baboon, rhesus macaque, and cynomolgus macaque indicating that the antibody is crossrcaotive with C5a from these species as well as with hum-m C>a.

The determination of funding affinity parameters to cynomolgus macaque ( fa was determined as described above. Briefly, the BNJ3tG antibody was screened against a-4 concentrations of recombinant eyriomolgus macaque C5a {antigen} using a capture technique as described above. The antibody was caponed by an Anti-Ec t human > directly immobilized on a (.'MS settsor chip vvith various concentrations in the ssug.e from 0 f nM to ?c> uM of cynomolgus C5a passed over the censor chip surface. The surface was regenerated with 2.0 rnM R{1, Ö 0.7% surfactant P20 (ft taco res after each cycle to sc-move bound antibody and antigen. The data was evaluated using Butane BIAcvsluation software using n f. 1 Langmuir Model Fit tRmaxtGlobai Fit. Ri'.Local Fit}. These experiments ware tb; screening purposes w ith a minimal number of analyte concentrations (3 to 4) with I duplicate. The tppros irmue Ks.? of the antibody tor eynomolgus macaque <35a is 3.3 nVI. .34 r fable 10 l<ibkJiL..Ain?uty,Dcremtjji<U!p Tea hrofems

This if- only an approximation ui the Kt> based on the quality of the curve fit.

The 8NJ383 antibody was also screened agamst 3-4 concentration* of recombinant mouse {.'5a (antigen) using a capture technique as described above to determine its affinity for mouse protein. The antibody was captured, «a described a bore, by an arui-Fc «human) directly immobilized on a CM5 sensor chip with various concentrations in the range from 0 h nM to 3 9 n.M of mouse CSa passed over the sensor chip surface. The surface was regenerated with 2() mM HCÈ. 0.02% P30 after each cycle ίο remove bound antibody and antigen. The data were evaluated using Blacorc BLA evaluation sett ware using a 1.1 Laugntuts Model Fit (Routs .Global Fit; R!: Local Fit). the above resells Indlealed tins· several of the humanized aolt-hCëa antibodies described herein arc e ro'-wrench a e with (35a Iforn set oral non-human primate species including cynomoigus macaque. rhesus macaque and baboon Fhc BN.mbh antibody. e.g, also erosxreacts w ith mouse (35a. Furthermore. die results described m this section nub cate that an ami-lnum-so ("5a antibody. such as BN J 383, Is useful not only hi clinical applications for treating complerncnt-assoetated disorders, bin also m a variety of pre-clnucal applications in non-human mammals, which are necessary for, or supportive of, approval of clinical use in human a Ϊ ^.;·.ι.νΛΠ :.i .S'.. .ti.:... 5:. iT.ViP.hÏVi /. >.=Λ. .¾: J.. Λ"?Λ /.l.sÜ /.1 ^. ΓΛ ?.. ΐ·. .ί.“.

An e.vperitnem was perlormed to evaluate -ho binding of an anu-( 5a amt body described herein. BN.I38.\ in the presence of potentially competitive amtgens Briefly, nuoeraimu labeled BN.ifxf <250 pM; v.ns incubated for loo hours id room temperature with i nM biotinylated 05a, alone with various concentrations te e., -loO, :η ). dd.-b i5X 1.6. am! 0.5 uMt of one of t he tbilooinn. ins human iff a uessui protein in phosphate-buffered saline, sbi human plasma, ιοί eynomosgns macaque plasma, id) Baip if 1mouses plasma, or (es DBA 2) {mouse) plasma With respect io the plasma component sbh (¾), {cl η ami {-:·), die concentration refers to the approximate final concentration of ("a a mi pen in tire incubation mixture.

Following the meubation period, the «samples were coru&amp;ctcd to respective individual wells of a sireptavidimcoated assay plate under conditions that allowed tor the binding of biotinylated idea to the streptavidm in the welts of 1 her (butte. The wells were washed those ugh iy to remove unbound material 'the amount of binding of BNjfbe to C5a in the presence of competitor was determined by detecting the a mo; mi of open tl produced from the detectable ruthenium label The results are shown m hg Λ>

Whereas human iff a dem’g was ,ut effective < vnspcttto;. there wa> virtually no eotnpetition observed m the pte^enee of mouse serum 17% induction m detectable suoo.fi observed at uppsoxintaiely a '400 i ratio oi Bale( mouse plasnat-deriv ed C5 to bunmylatcd hutuan ( 5a and 25% reductioti in detectable signal obsessed at approximately ,· *00:1 ratio of DB Af I phwna-dcrhed C5 to hiolmy feed human t '5ab No change m the level of binding ol BNIfff ίο biomsv Bled human ( fa was observed at up to appsoxmmiely a 15 1 ratio of human m cynomotgus masque p Banos-dem ed t. 5 to biotinylated btirnatr C5a.

As noted above, while the disclosure is m n»u way limited to any purrienlur theory or mechanism of acPon, the inventors hypo damme that the nrtfi-C.Ns antibody may bind to a subpopitbUon of tmeleevcd, processed if5 {com plasma ('5) eon.ohuinig less than ;0% oi the Total population of full length C5 in a sample <e.g„ a plasma sampled which subpopuladoo is in whole or in part denatured such that an otherwise occluded be a neoepttopc, to which the anti-05a anubody or fragment binds, is exposes). Thus, it is believed that the antibody docs not bind to a fully functional and or fully In net Sored ,species of C5 and thus docs not truly bind to uncleaved, native C.5. .Human plasma is at least an approximately 50 to 100-fold weaker compelitof for binding 10 biotinylated C'5a itetti fitffiii GSa dfsarg.

Notwithstanding these consideration!.', these results further indicate that ami-human CSa antibodies described herein, such as BNJ.'iH.h. preferentially bind to free human C5a ever; in the presence of up to approximately 2{Mold excess of uncleared, but not necessarily entirely native, plasma-derived human 05 protein.

Bkaihplê 16, Ifiset of anti< Sa ami bods- on AT and CF activity m vitm

An experiment was performed to evaluate the effect of an ant:-( 5a antibody described herein tBN.IeKV} on alternative pathway tAP) complement activity in viir« twine, pooled normal human serum iPNHSr The experiment utitired tbc Wieaksbw Alternative Pathway Complement Kit t Wiesktbw COMP!. ΛΡ350, Kuro-Diaonosiica, Sweden) and the associated protocol was followed w ith only routine optintHeoon welt w ithnt the purs tew of one of ordinary skill ht the art Bnctly aiiouofs of the PNH5 were sneubaied it! ’wells of u npopol>sacchnndewoateu plate tor one hour tat 5"·'°0ί alone w ith various ooneerurfmoos ίΟΗ?!··', 0.1)89, 0.194., 0.097, 0.049. 0.024. 0.012, 0.00A 0 004, and 0.002 μ M > of an awh-hi o antibody or an nmi-h05a antibody < BN ΠΧΐ) The a at I-('5 antibody inhibits the cleavage of human C'5 into fragments €5u and f 5 b, As a negative control., several wells were incubated whh PNHS under the same conditions, but hi the absence ofantt-hi'5 antibody or amj-h< 5u antibody.

Follow ing the incubation, the weds were washed thoroughly with kit-supplied IX was.lt buffer. The lose! of a hernat Is c pathway complement activation was measured by absorbance at 405 nm, follow mg contact of each well with a kit-supplied engynw conjugate!an anti~h'5b“9 antibody conjugated to alkaline phosphatase} ami fiuorogeme subsirate j which is operated upon by the cnxymet and incubation for 50 minutes at room temperature. The results are shown in Fig. U).

While the a.uti-C'5 antibody inhibited alternative pathway complement activity completely at concentrations greater than 0.1 μΜ the amt-hCfa antibody did not .significantly inhibit complement activity even at the highest concentration tested.

An cxpcnmem was per formed to evaluate the effect of an arm-Cda antibody described herein tBXidKb} on classical pathway t('P) con spiereen t activity b? r.<m; using PNRS. The experiment uiiiiacd the Wicsiabt*'· Classical Pathway Complement Kir {Wiesiab'Kt COMP!.. CP3 1Ö, Faro-Diagnoshoa, Sweden) and ihc associated protocol was followed wt?h only rot; in a; optimization well within the purview of the ordinarily-skilled artisan. Brief!) aliouots of the PNHS were incubated in wells of a human IgM antibody coated plate tor one hour tat 3rvCt along wtih various concentration.-* (7.2. 3.0, j X 0.9. 0,45, 0.2. 0,1,0.Ό5, 0.02, or 0.01 itM t of un ami-hid) antibody or an anti-hCSa antibody iBN,B83K s he anb-Ce antibody inhibits the cleavage of human Cé into fragments 05a and Oh. Λ* a control, sexcml wells were inctshated wtth PNHS urtder the same coed ebons, Intt tn the absence of attii-hCb antibody or arui-hC5s antibody.

Following the incubation. she wells w-ere washed thoroughly with kit-supplied IX wash butter. The lex el of alternative pathway compiemeru activation was measured by absorbance at 405 nm, following comad of each well with a kit-supplied enzyme conjugate tan «mi~C5f>9 antibody corns tasted to alkaline phosphatase) sad tluorogcnie substrate fw-hich is operated upon by the easy me; and incubation for 50 nanuses .at room temperature. The results use shown in Fig. 11.

While the sn?i-C5 enbhodx inhibited classical pathway complement activity completely at concentrations greater than 0.1 μΜ. the antt-hf éts antibody did not significantly inhibit complement »clt\ it> even at the highest concentration tested.

Takers together, these results Indicate that., w wuvxx the anti-hOa antibody, BNÜ383, did not significantly affect €51>b generation (terminal complement activation) driven by either the classical or altcmatixe pathway of complement, thus giving further evidence that the anti-hCSa antibodies described herein specifically latge? the free €5a adsjyhyiatoxhi arm of-Oompicmont sefiyafidh.. ï :.1 .. Λ. .T.:.. .Vi.t Λ . AMlVA.l^S :s.s!x. i.'.s' Aï'.M λ ï.^ . >Λλ1.. ίΛ .SÏ .S' .Μ. ΡΛ . .v: λίΑ Λ ϋίι'.ΤΛΓ. h; λ': .MV^. ΓΓΛ ii. i'ïn.sÏ. .ί'.ΟΛί .si Λ=Λ!?. !λ ί.. !.Vi

Ma&amp;ioQsua^ I he anti-CSa antibody BNJ3X3 (set tort!; ..shove) w ax utenbuted at --1 "C tor Wt hunt's hi the presence of a 2 1-fold molar escess of hurmm ('5 {him} to allow lor complete amibody;C5 complex formation. A parallel experiment o vs nudbrnted tomra an amibods that hi ruls ίο fra man €5 a; a 2:1 stoichiometry Baan: in a iter she a.m-;-f 5 antibody I Antibody :C 5 complexes w are resolved on a ISK iN! G-4000 $\V sixe exclusion column 1 Tosoh, Tokyo? UMug a Watess5''5 2(>vO;5 HFl.i' system w;ih a Waters*'5 \Y2~IK? dsud wavelength do; veto r to determine the occupancy of binding sites beaks 'aero monitored at a wavelength of 2 14 a no Ihe mobile phase for the HPLC analysis contained the follow mg budor comporahom 5.v mM laid POo.l rnM Nag 1 PC w, and I 50 mV! NaCL at nH'wl Use How rate was 1.0 ml per mi η me and she run time was 20 smtunes Dau were uecjmred and analysed with Waters fimpowerrN1 2 chromatography λοΙολ are

As depicted ht 1- Ip- 12, ENi )83 alone (Fig, :2;V? and hC5 alone (Fig. 12B> each resolve as a single peak centered around 10.2 mimues tmd w ΥΝΝ of the a no--('ha antibody thC’5 so mo I es.es resolve as a single peak ccnfesed a round 9.2 minutes t figure 12C). In con mast, complexes ofhCS and dtc arnn-C'o antibody resolve in two peaks centered at 8.6 min < 3:->%? and 9 2 ram in IGd (Fig. 12D! Thus, even m a molar ex raws 0 t' Μ to 95.2% of lira BN j.Wt has a tree boh arm that may be capable of buiding to human 05a.

These samples were further examined to determine if the BNJ >83 antibody retained the ability to bind 05a in the presence of saturating concentrations of 05. Free antibody or am foody :€$ complexes were t itrated from 50*} to 0.5 og/mL on a sirernavidin craned plate to which biotin conjugated hC5a rans menu 4% Iced. Captured antibody vwra detected w ith an nnii-human Fc antibody conjugated to horse radish peroxidase, The results depicted In Fig. 13 demonstrate that even BN j 383 completed with hC5 is capable 01 binding C 5a and that the concentration of antibody available to bind ('Sa is not deteeuhk diminished in the presence of saturating C5. Thus, ihe results described herein indicate that the antibody, oven ht the presence of a molar excess of tmeleaved Cm retains the nbihiy to bifid to five C5« with hU/h aUbuty and diere by retain . exen is- that molar atmess, tie sbiilty to IxsMifeit activity oiMfSa, ^ 4 aeons et o t?.CAMdMLconMM^

Oxmonsosgus macaques vore administered mtnwenonsh s siftgte dose of she 8NJ3H3 anh-Cêu ana body m 1 mg'kg, 10 mg-'kg. 100 mg;kg 250mg/kgor -100 mg'kg. Plasma samples were collected frorri the macaques at time point'» ranging hom I day to 30 day1·; tol km mg the admimstrabon of the antibody, Levels of Cda'idd.a desorg in plasma were determined by an decirochomihuninesecnt tECij assay in which a Boo (!5a Cent demure was eapiarcd ou a microliter plate coaled witlt an antibody specific for a neoepltopeon (" 5aC5,t dosurg and detected with a non-competitive b oa antibody conjugated to a rnihenate containing EC L moiety and read on a SECTOR 24ϋΟ;Μ plate reader (McsoSculc Discover}'}. Circulating amibods undent rations wore determined by and enzyme Imbed immunosorbent assay {ELISA} m which free antibody tBNi IMM νγ«s captured on a microliter plate coated with human C5a. denary and delected with » mouse a me human antibody conjugated ?o horseradish peroxidase < BRP). ds shown in Fig. 14. like rise results described in Example 13, circulating concentrations ei'BNj.ifa as- low as lb isg.Tnl.. deplete plasma C5u C5a desarg levels to below detectable limits in cynomoigus monkeys. These results also underscore that the antibody, even in the presence of a molar excess of uncleuved CS.. retains the ability to bind to free C5a with high affinity and thereby retains, even in that molar excess. !her ability to inhibit the pro-irbiammasoty activity of Com

To determine whether BN.M85 had an effect on hemolytic activity of macaque scrum, the antibody wan evaluated hi an m vitm red blood cell hemolysis assay.

The red Mood cell hemolysis assay is generally described in detail in. e g.. binder et al i 14451,/ C 7 in Inw w 4o; 15Mb I MM. Brief Is scrum samples obtained trom macaques administered BNJ5K3 {us described above) were added to rtmlaple weds of a °b web assay plate such tltat the eoneettiration ot the serum in each wed was approximate!} 10"... Ihe -wrum samples, bx virtue of the time points nt w hich they were obi a meek, contained various otntcenlratious of the BBJ.M r antibody. The hemolytic acitvity of serum from macaques not roccK iiiy BNBkd sens'd a* negative control and as diC: baseline hemolytic activity kveh

Oneken erythrocytes <l.or«piro Biological Laboratories, Piporvillc. PA) were o ashed und resuspended m buffer at a final concentration of.'? x 10 cells/mL The erythrocytes were sensitised to ly^is· by incubating the cell·.·» with a?* anti-chicken red blood eel) polyclonal antibod)· composition. The aeushked erythrocytes were added to the wells of the 96 well plate and the plate was militated at 37''C' tor TO minutes. Hemoglobin release sv«s measured by apparent absorbance ;u 4 i S nrn using a rmo opiate reader;..

As shown in Fig. 15Λ. even high concent radons of BN.I333 did not substantially inhibit erythrocyte hemolysis under these ax vivo hemolytic assay experimental conditions

The BNJ383 antibod) was also evaluated to determine if it had ae effect on complement activation of macaque serum using an c.v vivo OHSOeq assay. T he C'HiOcq essay is a method for measuring the total classical complement activity in serum t his test is an ermyrue linked smrnunosorbent assay, winch uses human gammatilobulins and otou.se monoclonal antibodies as the activator of the classical complement nathnay and captures the terminal complement complex (TCO generated on a rmermite? well coated with a I CC ttetsepitope specific antibody. Captured TCC' is detected w uT? a goat uriii-TCC antibody conjugated to horse radish peroxidase. The C H50e<q assay provides a direet measure of terminal complement complex fj'CO formation

As shown in Fig. 15.B. high concentrations of BNBta present it? the macaque scrum were capable of substantially inhibiting TOC formation under these cv mv, conditions, irtusc results indicate that the BN.I333 antibody is not only capable of binding to and sequestering h e·-.. Cëa but is also capable of. as a function of concentration, partially or substantially inhibiting TOC formation.

An or vivo expertmeft! was also performed to evaluate the effect ofBNBk.il on classical pathway {CP? complement activity using the macaque serum samples described above The' experiment utilised the WiesinfhH Classical Pathway Complement Kit (Wiesialrte COMPL OPa Μ), ΈmrwDi agnoxtiea, Sweden1 and the associated protocol was followed wuft only routine optimization w cl I within the puss ww oi the ordinarily-skilled artisan. Brietly aliquots of Ha.' macaque serum samples warn meuhared In wells of a Inimar: tgM antibody-coated plan: lor one four. As a control. seven) wells were ira;.ubreed under the seme conditions w uh serum from macaques not administered the RNI3h3 antibody.

Following da: incubation, die webs were w ashed thoroughly w ith kit-supplied I X wa-dt belie;. The He-el ol ahernuth e pathway complement activation was measured ip absorbance at -105 nm. following contact of each well with a ktf-suppSiecl enzyme conjugate tan anti-OM) antibody conjugated to alkaline phosphatase} and iTtosogcnu: substrate (which is operated upon by the eretyurc t and incubation for 30 minutes at room temperature The results are .shown ht hie. I5C

The ann-hCda antibody did significantly. though not completely, inhibit complement activity in a dose-dependent manner. Taken together, the results described hetein indicate that 8NJ 3db is not only a potent antagonist off 5 a, hut i- a Iso an incomplete partial antagouna of terminal complement complex formation >? who. .Huts, the anlibody and unPbodtes sharing, its properties are useful ibr treating a variety of complement-associated disorders m whtclt Ccu-medlatcd ittHatumatioa is the primary contributor to deleterious pathological effects and ICC may play a less significant or even beneficial role in the pathology.

Whtie dte present disclosure has been described w uh reference to the specific embodiments thereof, it should be understood by those skilled in the an that various changes may be made ami equivalents may be substituted without departing from the true spirit and scope of the disclosure. In addition, many nutdificutions may be made to adapt a particular mmahon. material., composition of mutter, process, process step tar steps, so the objective, sptnt and scope of the ruvseni disclosure. .Ml such modifications are intended to be within the scope of the disclosure.

Claims (16)

1. What is claimed is

   1. An isolated antibody, or antigen-binding fragment thereof, that binds to a free C5a polypeptide, comprising a light chain CDR1 comprising the amino acid sequence depicted in SEQ ID NO:20, a light chain CDR2 comprising the amino acid sequence depicted in SEQ ID NO:21, a light chain CDR3 comprising the amino acid sequence depicted in SEQ ID NO:22, a heavy chain CDR1 comprising the amino acid sequence depicted in SEQ ID NO:28, a heavy chain CDR2 comprising the amino acid sequence depicted in SEQ ID NO:46, and a heavy chain CDR3 comprising the amino acid sequence depicted in SEQ ID NO:47.
2. 2. The isolated antibody, or antigen-binding fragment thereof, of claim 1, wherein the antibody, or antigen-binding fragment thereof, comprises a light chain polypeptide comprising the amino acid sequence depicted in SEQ ID NO:42 and a heavy chain polypeptide comprising the amino acid sequence depicted in SEQ ID NO:45.
3. 3. An isolated antibody, or antigen-binding fragment thereof, that binds to a free C5a polypeptide, comprising a light chain polypeptide comprising the amino acid sequence depicted in SEQ ID NO:42 and a heavy chain polypeptide comprising the amino acid sequence depicted in SEQ ID NO:45.
4. 4. The isolated antibody, or antigen-binding fragment, of any one of claims 1 to 3, wherein the antibody, or antigen-binding fragment, is selected from the group consisting of a monoclonal antibody, a humanized antibody, a fully human antibody, a recombinant antibody, a bispecific antibody, a single chain antibody, a diabody, an intrabody, a chimerized or chimeric antibody, a deimmunized human antibody, an Fv fragment, an Fd fragment, an Fab fragment, an Fab’ fragment, and an F(ab’)2 fragment.
5. 5. The isolated antibody, or antigen-binding fragment, of any one of claims 1 to 4, wherein the antibody, or antigen-binding fragment, binds to C5a with a KD that is less than 7.5 x 10"11 M.
6. 6. An isolated antibody, or antigen-binding fragment thereof, that crossblocks the binding of the antibody, or antigen-binding fragment thereof, of any one of claims 1 to 3 to C5a.
7. 7. A pharmaceutical composition comprising the antibody, or antigen-binding fragment thereof, of any one of claims 1 to 6 and a pharmaceutically-acceptable carrier.
8. 8. A method of treating a human afflicted with a C5a-associated complement disorder, said method comprising administering to the human the antibody, or antigen-binding fragment thereof, of any one of claims 1 to 6.
9. 9. The method according to claim 8, wherein: (i) at least 1 mg of the antibody, or antigen-binding fragment thereof, per kg body weight of the human is administered to the human to thereby partially or completely bind and sequester nanogram levels of free C5a for at least 12 days, or (ii) the antibody, or antigen-binding fragment, is administered to the human in an amount sufficient to (a) achieve molar Cmax values substantially lower than the molar physiologic concentration of uncleaved hC5 and (b) partially or completely bind and sequester pathophysiologic levels of free C5a.
10. 10. The method according to claim 8 or claim 9, wherein the C5a-associated complement disorder is selected from the group consisting of atypical hemolytic uremic syndrome, age-related macular degeneration, severe burn, rheumatoid arthritis, sepsis, lupus nephritis, a complement-associated pulmonary disorder, and antiphospholipid syndrome.
11. 11. The method according to claim 10, wherein the complement-associated pulmonary disorder is asthma or chronic obstructive pulmonary disease.
12. 12. The method according to any one of claims 8 to 11, wherein the antibody, or antigenbinding fragment thereof, is administered to the human in an amount and with a frequency effective to maintain a reduced level of systemic C5a activity for the duration of the treatment.
13. 13. The method according to any one of claims 8 to 12, further comprising, after the administering, monitoring the human for an improvement in one or more symptoms of the C5a-associated complement disorder.
14. 14. The method according to any one of claims 8 to 13, further comprising administering to the human one or more additional therapeutic agents.
15. 15. The method according to claim 14, wherein the antibody, or antigen-binding fragment thereof, is delivered to the human by intravenous administration, intrapulmonary administration, intraocular administration, subcutaneous administration, or intraarticular administration.
16. 16. Use of an antibody, or antigen-binding fragment thereof, of any one of claims 1 to 6 in the manufacture of a medicament for the treatment of a C5a-associated complement disorder.