TW416960B - Reshaped human antibody to human interleukin-8 - Google Patents

Abstract

Reshaped antibody to human interleukin-8, comprising; (A) L-chains comprising; (1) human L-chain C-region, and (2) L chain V-region comprising human L-chain FRs and L-chain CDRs of mouse monoclonal antibody to human IL-8; and (B) H-chains comprising; (1) human L-chain C-region, and (2) H-chain V-region comprising human H-chain FRs and H-chain CDRs of mouse monoclonal antibody to human IL-8. Since major parts of this reshaped human antibody are derived from human antibodies, and CDRs have low antigenicity, then the present reshaped human antibody has a low antigenicity to human, and therefore is promising for medical use.

Description

A7 B7 ί * '4 1 6960 V. Description of the invention (1) The present invention relates to the complementarity determining region (CDR) and variable region (V region) of a mouse monoclonal antibody against human interleukin-8, and Anti-human interleukin-8 human / mouse chimeric antibody, human light chain (L chain) variable evidence and human heavy chain (H chain) variable M complementarity determining region (CDR) are anti-human Reconstituted human antibody replaced by CDRs of a mouse monoclonal antibody to Leukin-8. The present invention further provides DNA that can encode the above antibody, or a part thereof. The present invention also relates to a vector formed by containing the DNA, particularly a performance vector, and a host transformed by the vector. The invention also provides a method for manufacturing a chimeric antibody against human interleukin-8, and a method for manufacturing a reconstituted human antibody against human IL-8. Anti-human interleukin ~ 8 (IL-8) was found from the supernatant of monocyte culture stimulated with lipopolysaccharide (LPS), and is called Monocyte-derived neutrophil Cheinotactic factor (

Ji D N C F) or n e u t r o p h i 1 a c t i v a t i n g p r o t e i η-1 (N A P-1) and other mobile hemoglobin (Chemokine). The IL-8 series can be produced from various cells printed by the I-Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. It acts on polymorphonuclear leukocytes and lymphocytes, and has the activity to make it move according to its concentration gradient (Chemotaxis). In addition, it not only induces the movement of neutrophils, but also has the functions of activating granulocytes by releasing granulation, releasing active oxygen, and adhering to endothelial cells. In inflammatory diseases, more specifically, in cystic pulmonary fibrosis, idiopathic pulmonary fibrosis, adult tachypnea syndrome, sarcoidosis (sarCGidQsis), respiratory diseases such as purulent pleurisy, and skin diseases such as psoriasis, And chronic rheumatoid arthritis, local ileitis, ulcerative colitis. The paper size is applicable to Chinese National Standard (CNS) A4 (210X297 mm) Α7 Ύ 4 1 6960 _'_ B7__ ___ V. Description of the invention (2 ) And other diseases of these diseased nest sites can be seen pathologically with white blood cell infiltration. In addition, IL-8 was detected in test substances from patients with these diseases, and it is speculated that it plays a very important role in inflammation. (McElvaney, NG, etc., J. Clin. Invest., 90, 1296-1301, 1992, Lynch III, JP, etc., Am. Rev. Respir Dis., 1 4 5, 1 4 3 3-1 439, 1 9 9 2. Donnelly, SC, etc., Lancet (3 4 1, 6 4 3-6 4 7, 1 9 9 3, Car, BD., Etc., A π. J Respir. Crit. Care Med., 149, 655- 659, 1994s * Antony, V. B., etc., J. I mmu η o 1., 1 5 1, 7 2 1 6-7 2 2 3, 1 993, Take matsu, H., etc., Arch · Dermato 1 ., 1 2 9, 7 4 -8 0, 1 9 9 3, B re η na π, F · M. Etc., E ur. J. I id mu η o 1 ·, 2 0, 2 1 4 1- 2 1 4 4, 1 9 9 0, I zz., R · S · etc., S cand. J. G as I: roentero 1, 2 8, 2 9 6-3 0 0, 1 9 9 3, I zzo Ιϊ .S, et al. Am. J, Gastroenterol., 87, 1447-1452, 1 9 9 2) 0

Ko, Y, C. and other systems have prepared human IL-8 as an antigen to immunize mice, thereby binding to human IL-8, thereby binding to inhibit human IL-8 binding to neutrophils. That is, it can neutralize humans. The Central Standards Bureau of the Ministry of Economic Affairs: H Consumer Cooperative printed the biologically active mouse monoclonal antibody WS-4 of IL-8 :. The same type of mouse monoclonal antibody WS-4 is known as the K-type L chain and the C r 1 plastic chain (J. Immunoi. Methods, 149, 227-235, 1992). Known anti-human IL-8 antibody systems other than WS-4 are A. 5.2.1.14 (Boylan, A.M. et al., J. Clin. Invest., 89, 1257-1267, 1992), International patent application paper size applies to Chinese national standards (CNS &gt; A4 size (2 丨 0X2.97 mm) t '4 1 6960 A7 B7 V. Description of invention (3) Please W09 2-0 4 3 7 2 Revealed anti-Pep-1 antibody or anti-Pep-3 antibody or DM / C 7 (Mulligan, MS, etc .... Immunol, 150, 5585-5559, 1993) etc. 0 In the experimental system using rabbits, Administration of mouse monoclonal antibody w S _ 4 in pulmonary ischemia and reperfusion disorders (Sekido, N. et al. Nature, 3 65, 654 — 657, 1993), 1 ^ 153 induced dermatitis (H arada, A, et al. Nterna t1. Immunol., 5, 681-690, 1993), LPS or interleukin-1 (IL-l) -induced arthritis ('Akahoshi, T. et al. Lymphokine and Cytokine Res., 13, 113-116, 1994) found that neutrophil infiltration has been inhibited. The human IL-8 homologue (homologue) also exists in rabbits printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs, which is called .1 L-8. It is known that mouse monoclonal antibody WS-4 line cross-reacts with rabbit IL-8 and can inhibit rabbit IL-8 binding to rabbit neutrophils (Harada, A. et al., Internatl. Immunol ., 5 »681-69 0-1993), so it means that as a therapeutic agent for the treatment of inflammatory diseases in humans, the anti-human IL-8 resistance system is extremely useful. A single-antibody system from mammals other than humans Humans have a high degree of immunogenicity (sometimes called antigenicity). For this reason, the medical treatment of humans is limited. For example, when mouse antibodies are administered to humans, they will also become impurities and foreign substances and will be metabolized. The half-life is relatively short in humans, and its effects cannot be fully exerted. In addition, the Chinese national standard (CNS) A4 specification (210X297 mm) is applicable to the human paper size of the mouse antibody administered: 4 1 6960 A7 B7 Printed by the Consumers ’Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs • Invention Description (4) Anti-mouse antibodies often cause * 3 seropathies or other allergic reactions, etc. Good and dangerous immune response 0 Based on this 9 Jar, Jing? Method of administering mouse antibodies to humans several times. To solve these problems, a method of manufacturing humanized antibodies has been developed. 0 Mouse antibodies can be used in two ways. The simpler method is to make a variable region (V region) for the original mouse antibody and a fixed region (C region) for the chimeric antibody of a suitable human anti-Λάϋ body. 0 The resulting chimeric system It originally contained the full variable region of mouse antibodies, so it can be expected that it has the same specificity as mouse antibodies and binds to the antigen. The chimeric anti-WUh body has a protein sequence ratio other than that of white people. It has been substantially reduced, so it is predicted that the immunogenicity will be lower than that of the mouse antibody. Although the chimeric antibody can fully <cL · 7m bind to the antigen, its immunogenicity is low. 9 It is possible that it may also generate an immune response to the variable region of mice (L 〇Bu g 1 i 〇) A. F, etc. Λ Pr 0 C Na th Ac ad-Sc i. USA) 8 8 4 2 2 0 — 4 2 2 4 9 1 9 8 9) 0 The second method of humanized mouse antibodies is more complicated, but it can significantly reduce the potential immunogenicity of mouse antibodies. 0 In this method, the white mouse antibodies are variable. Τ only transplants its complementarity decision (Co mp 1 e me ntar it determ in g re gi on CDR) to the human variable region to make "reshaped" human antibody variable region. Make the reconstructed CDR structure of human variable Tg? Closer to the original structure of mouse antibodies. Sometimes, the mouse antibody can be transplanted to support the CDR framework region (FR)-part of the protein sequence into the human variable region. 0 Secondly, this reconstructed human variable region is connected to the human fixed region. 0 Paper size: General Chinese National Standard (CNS) A4 specification (210X2.5 »7mm) f Γ 416960 Α7 _ Β7 V. Description of the invention (5) The part of the reconstructed human antibody from the human protein sequence is CDR And very few of them (FR). The CDRs are composed of hypervariable protein sequences. These do not have species-specific sequences. For these reasons, reconstituted human antibodies carrying mouse CDR should no longer be more immunogenic than natural human antibodies containing human CDRs. Reconstructed human antibodies can also refer to Riechma η η, L. et al., Nature, 332, 323-327, 1988; Verhoeyen, M. et al., Science, 239, 1534-1536, 1988; Kett1eborough, C. A. et al., P rotei η E π gng ·, 4, 7 7 3-7 8 3, 1 9 9 1; Maeda, H. Et al., Human Antibodies and H ybridoma, 2, 1 2 4-1 3 4, 1991; Gorman , SD et al., Proc. Natl. Acad. Sci. USA, 88, 418 and 4185, 1991; Tempest, PR et al., Bio / Technology, 9, 266-271, 1991; Co, MS et al., Proc.

Natl. Acad. Sci. USA, 88, 2869-2873, 1 99 1; Carter, P., etc., Proc. Natl. Acad. Sci. USA, 89, 4 285-4 289, 1 992; Co, MS, etc., J. Immunol ,, 1 4 8, 1 1 49 '1154, 119 2; and Sato, K., etc., Cancer Res., 5 3, 8 5 1-8 5 6 Printed by the Consumer Cooperative 1 993 ° As mentioned above, reconstructing the human anti-system is extremely useful for treatment according to the theory, but the reconstructed human antibody against human IL-8 is not known. In addition, the method for rebuilding human antibodies cannot be applied to methods generally applicable to any specific antibody. Therefore, in order to produce reconstructed human antibodies that have sufficient binding activity or neutralizing activity for specific antigens, it must take a lot of time (such as Sato, K., Cancer, Res. 53, 1-6 (1993). This paper is applicable to China Standard (CNS &gt; A4 specification (2iOX297mm) _ β _ printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs f * 41 6960 V. Description of the invention (6)). The present invention can provide anti-human i L- Reconstructed human antibody of 8. In addition, the present invention can also provide a human / mouse chimeric antibody that is extremely useful in the process of making the reconstructed human antibody. The present invention can further provide fragments of reconstructed human antibodies. The expression used for manufacturing chimeric antibodies, reconstructing human antibodies and these fragments is provided. The present invention further provides a chimeric antibody against human IL-8 and a method for manufacturing these fragments, and a reconstructed human antibody against human IL-8 and the like. Method for producing fragments of the same. More specifically, the present invention provides: (1) the L chain V region of the mouse monoclonal antibody against human IL-8; and (2) the anti-human monoclonal antibody against human IL-8 Chain V region The invention further provides: (1) an L chain formed by a human antibody L chain C region and an L chain V region of a mouse monoclonal antibody against human IL-8: and (2) a human antibody limulus chain C region, and The Η chain formed by the Η chain V region of a mouse monoclonal antibody against human IL-8, the present invention further provides: (1) a mouse chain containing a human antibody L chain C region, and a mouse monoclonal against human IL-8 The L chain formed by the L chain V region of the antibody: and (2) the V chain formed by the V chain of the human antibody Η chain C1E and the anti-human IL-8 mouse monoclonal antibody: the formed anti-human IL Chimeric antibody to -8. The present invention also provides: (1) the L chain V region of the mouse monoclonal antibody WS-4 against human IL-8; and the standard of this paper applies the Chinese National Standard (CNS) A4 specification ( 210 &gt; &lt; 297 mm) —〇_ (Please read the precautions on the back of the book before filling in this page)

I * 4 1 6960 A7 B7 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs. 5. Description of the invention (7) (2) Anti-human IL-8 anti-mouse monoclonal antibody WS-4 4 chain V region. Provide (1) the L chain containing the L chain V region of the human monoclonal antibody WS-4 and the human L chain C region and anti-human IL-8 and (2) the C chain region containing the human limulus chain and the anti-human IL-8 The mouse monoclonal antibody WS-4 is formed by the Η chain V surface of the Η chain. The present invention also provides a mouse monoclonal antibody WS-4 which contains (1) a human L chain C region and anti-human IL-8. The L chain formed by the chain V region and (2) a mouse monoclonal antibody WS-4 containing human C chain and the anti-human IL-8 chain including the anti-human IL-8 chain formed by the Η chain formed by the chain V region Chimeric antibodies (1) CDRs of the anti-human IL-8 monoclonal antibody L chain C, and (2) CDRs of the anti-human IL-8 monoclonal antibody Η bond V 15? The present invention also provides (1) anti-human I CDRs of B-8 mouse monoclonal antibody L chain V; and (2) CDRs of mouse monoclonal antibody anti-human IL-8 monoclonal antibody Η chain V region. The present invention also provides the paper standard applicable to Chinese national standards (CNS ) A4 size (210X297mm) -10 Please read the notes on the back again 'The Central Ministry of Economic Affairs is the printed bag of negative consumer cooperatives at Γ 416960 Β7 5. Description of the invention (8) Contains (1) human L chain V Frame structure region (FR); and (2) a reconstructed human l-chain V region of an anti-human IL-8 antibody formed by the CDRs of the l-chain V region of a mouse monoclonal antibody against human IL-8; and (1) FR of human Η chain V region: and (2) anti-human IL-8 mouse monoclonal antibody Η chain V region C Reconstitution of human anti-human IL-S by DR to rebuild human V chain. The present invention further provides: (1) a human L chain C region; and (2) a human L chain FR, and an anti-human IL-8 mouse monoclonal antibody L chain CDR formed by an L chain V region; Reconstruction of human L-8 antibody against human L chain; and (1) human C chain containing C region; and (2) human CDR chain containing FR, and mouse monoclonal antibody against human IL-8 Chain V region: Reconstructed human Η chain formed by anti-human IL-8 antibody. The present invention also provides a reconstituted human antibody against human IL-8, which is: (△) a mouse containing (1) human 1 chain 0 region, and (2) human L chain FR, and anti-human IL-8 L chain formed by monoclonal antibody L chain and CD R: and (B) formed by (1) human 单 chain C region, and (2) human FR, and anti-human IL-8 mouse monoclonal antibody Η chain CDR Η chain: made. More specifically, the present invention can provide the paper size applicable to the Chinese National Standard (CNS) A4 specification (210 X 47mm) _Gongong (Read the notes on the back of the poem before filling this page)

l · 416960 B7 Printed by the Central Consumer Council of the Ministry of Economic Affairs, Employees' Cooperatives. V. Invention Description (9) 1 (1) Anti-human IL-8 anti-mouse monoclonal antibody ns. IWS-4 L chain V region shown in the following sequence CD R Ο t 1 CDR 1 t A rg A 1 a S er G 1 U 11 e 11 e Ty Γ S er Ty r L eu 1 I 谙 i first 1 A ί a read I 1 1 CDR 2 As η Ala L ys Th Γ Le u A 1 a As ρ back 1 I 1 CDR 3; G 1 η H is His Ph e G 1 y Ph e Pr 0 Ar g Th r Note i 1 gives 1 and the term is again, 1 隹 (( 2) Mouse monoclonal antibody against human IL-8 shown in the following sequence: 1 WS — 4 CD R of the Η chain V region, 1 CDR 1; As P Ty r Ty r L. eu Se r! 1 CDR 2 Le UI! E A rg As η Ly s A 1 a As η G 1 y Ty Γ Th rj! Ar g GI u Ty r Se r A 1 a Se r Va 1 Ly SG iy Order | CDR 3 G 1 UA sn T yr Ar g Ty r As P Va 1 G 1 U Le UA 1 a 1 [Ty r 1 I and 5 The present invention provides 1 1 frame structure region (FR) containing (1) the human L chain V region: and iy (2) an anti-human IL-8 mouse monoclonal antibody W s-4 L chain Reconstruction of the anti-human IL-8 antibody formed by the CDRs of the V 1 1 region The human L chain V 1 1 I region and J -1 (1) FR containing the V region of the human Η chain: and (2) anti-human ϊ L 丨— 8 mouse CDRs of WS 4 H chain V region; anti-1 1 human IL — δ reconstructed human H chain V region 0 1 I The present invention also provides: 1 I containing {1) human L chain Zone C and (2) with human L chain FR 9 1 1 1 This paper size is applicable to Chinese National Standard (CNS_) grid (210X297 mm) _ 12

A7 B7 1 '41 6960 Printed by the Central Laboratories of the Ministry of Economic Affairs and Consumer Cooperatives. V. Description of invention (10) and anti-human IL-8 mouse monoclonal antibody WS-4 L chain CD R. ; Reconstructed human L chain of the anti-human IL-8 antibody: and (1) the human Η chain C region; and (2) the human Η chain FR, and anti-human IL-8 mouse monoclonal antibody WS — 4 The Η chain V region formed by the Η chain CDR; the reconstructed human Η chain formed by the anti-human IL-8 antibody. The present invention also provides a reconstituted human antibody against human IL-8, which is: (A) a mouse monoclonal antibody containing (1) a human L chain C region, and (2) a human L chain FR, and an anti-human IL-8 L chain formed by the L chain CDRs of the antibody WS-4; and (B) a mouse monoclonal antibody WS-4 containing (1) the human Η chain C region, and (2) human FR, and anti-human IL-8 The Η chain formed by the CDR chain CDR;--The present invention relates to polypeptides constituting the above-mentioned various antibodies, and DNAs capable of encoding the same. In addition, the present invention relates to a performance vector containing the above-mentioned DNA, The host of transformation. The present invention further provides a method for manufacturing a chimeric antibody and a fragment thereof against human IL-8, and a method for manufacturing a reconstituted human antibody and a fragment thereof against human IL-8. Brief description of the drawings: Figure 1 shows the expression quality of the human elongation factor la (Hl — a) promoter / enhancer, which is very useful in expressing the L chain and Η chain, respectively. This paper is based on Chinese national standards ( CNS) A4 specification (210X297 mm) (Please read the notes on the back before Vk page) Order, A .. -13-I1 41 6960 Amendment to Annex 1 of the Republic of China No. 83110990 Patent Specification Amendment Page A7 Β7

V. Description of the invention (11) Body Η EF — VL Biyue-g — k .7 · * · li'F-V Η-gr The second picture is to confirm the embedding of the present invention in cos cell culture supernatant The binding activity of WS-4 antibody (c hL / c hH) to humans is shown by the results of ELISA (enzyme-linked immu η osorbe η 1: a $ say). Invented the first version "a" (RVHa) of the human antibody Η chain V region, and the first version "4" of the WS-4-like antibody L chain V region "4" (DNA diagram of each amino acid sequence of RV. Figure 4 The reconstituted human WS-4 antibody (RVL a) and V chain V region (RVHa) of the present invention respectively represent the WS-4 antibody Η chain V region (c hH) and the chimeric WS L chain V region (chL) and COS Cells, compared with the chimeric WS-4 antibody (chc hH) of the present invention produced in COS fine solution, compared the binding activity to human IL-8 and the results of EL I SA production.

Figure 5 shows the results of EL I SA. Comparing the L chain V region of CIL-8 WS-4 and reconstituted human L a) is now L / amount on chimeric-4 antibody cell culture, with 0 S cell I- ------------------ Order --------- line ^ (Please read the precautions on the back before filling out this page) Employees of the Intellectual Property Bureau of the Ministry of Economy Consumption Cooperative printed L Ming, \} h hair / cah C this a H LH contains LVVV somatic VR RR anti-production R \, \ 4 cae a _ middle body LHL s liquid anti vvv w Cheng 4 RRR * close I, \ ， The figure embedding sb 'a amount Mingpei WH LH germinal cell type V vv and the human RRR sexual property s built \, \ Live 0 heavy ada joint c LHL knots and eight vvv8 liquid, into rrrI Cheng}, “\, Haaa f I raised on L, LHLH culture c VVV &gt; Renzhi / RR RR applies the national standard (CNS) A4 specification (210 x297 mm) for this paper size -14 — Ministry of Economic Affairs Printed by the Intellectual Property Bureau's Consumer Cooperatives _ rr 4ί6 ^ 〇I ^ ^ ~: ·. ： .. f ------- 5. Description of the invention (12) j8aa5! 6 l_i_ _nr — «· ——-- · &Quot; —— ·· 11 · 1 The 6th figure is expressed by the result of ELISA , Compare the chimeric WS-4 antibody (C h L / ch Η) produced in the culture supernatant of C, 0 S cells with the invention produced in the culture supernatant of C 0 S cells &gt; * Eight Reconstituted Human WS-4 Antibodies from RVLb (73 ^ 5 / '~ RVHa, RVLb / RVHb, RVLb / RVHc, RVLb / RVHd, gVLb / RVHe, RVLb / RVHf &gt; RVLb / RVHg &gt; RVLb / RVHh) Graph of human IL-8 binding activity versus yield. v Figure 7 shows the result of EL I SA, comparing the purified chimeric WS-4 antibody (chL / chH) of the present invention with the purified reconstituted human WS-4 antibody RVL a / RVHg and RVL b / RVHg of the present invention. Binding activity profile for human IL-8. The details are as follows.

The DNA of the mouse V region can be selected as the gene encoding the mouse monoclonal antibody V1S of anti-human IL-8, and it is necessary to make a synthetic tumor that can produce anti-human IL-8 mouse monoclonal antibody. As the source of the gene. After the mRNA is extracted from the fusion tumor, it is changed to single-stranded c DNA according to a known method, and the target DMA can be prepared by polymerase chain reaction (PCR) method. The source of this gene can be a fusion tumor WS_4 produced by Ko, Y. C., etc., which can produce a monoclonal antibody against human IL_8 mouse. This production method has been described in J. Immunol. Methods, 149 * 227—235 * 1992, and it is described in Reference Example 1 ° (1) Full RNA is adopted-This paper standard applies Chinese National Standard (CNS) 8-4 specifications （2IOX297mm） (#Please read the notes on the back before filling in this page)

at X 416960 B7_ V. Description of the invention (13) The target DNA is purified to encode the V region of the mouse monoclonal antibody. The fusion tumor is destroyed by guanidine alanine treatment, and the density gradient centrifugation (Chrgwin, JM) is performed. Et al., Biochemistry, 18 &gt; 5 2 9 4 — 5299 * 1979), and the whole RNA can be obtained. However, it is also possible to use the methods that have been used when selecting genes for other proteins, such as surfactant treatment in the presence of ribonuclease inhibitors such as vanadium complexes and phenol treatments (Berger, SL, etc. Bioehemistry,! · 8 , 5143-5149, 1979) °

(2) Synthesis of c DNA Secondly, the oligo (d T), which is locally present at the 3 terminal end and is complementary to the poly A chain, is used as a primer, and the mRNA containing the entire RNA obtained as described above is used as a template Single-stranded cDNA complementary to RNA synthesized by reverse transcriptase treatment (Larrik, JW et al., Bio /

Te chno 1 ogy, 1,934 ~ 938 1989) ° Also, random primers can be used at this time. In addition, if you only want to obtain mRNA, you can treat the whole RNA with oligo dT weaver layer column and isolate only the .mRNA with polyA chain. Printed by the Central Laboratories of the Ministry of Economic Affairs and Consumer Cooperatives (谙 Please read the notes on the back before filling this page) (3) Amplify the DNA encoding the V region by polymerase chain reaction ° Second, use the polymerase chain reaction ( PCR) method to specifically amplify the c DNA encoding the V region. To amplify the carba (k) -type L chain V region of a mouse monoclonal antibody, 11 types of oligonucleotide primers (Museuse K appa V ariab 1 e; MV) shown in the sequence 1 to 11 were used, respectively. And sequence 1 2 τπ oligomeric succinic acid (Mouse Kappa Constant 丨 MKX) as the paper standard is applicable to the national standard 〇 (〇 ^) 8 4 specifications (210 乂 297 mm> _16_ Ministry of Economic Affairs Central Standards Bureau off-load consumption Cooperative printed Γ 416960 A7, B7 V. Description of the invention (14) is 5 &lt; terminal primer and 3 V terminal primer. The above MKV primer system is fused with a DNA sequence encoding a mouse carba type L chain leader sequence, and the above MKC The primers are fused to a DNA sequence that encodes the mouse Caba type L chain C region. To amplify the Η chain V region of a mouse monoclonal antibody, 12 oligonucleotide primers shown in the sequence 1 3 to 24 are used (Mouse Heavy Var i abl e; MHV) and the oligonucleotide primers (Mouse He a. Vy Constant; MHC) shown in Sequence 25 are used as 5 'end primers and 3 ^ terminal primers, respectively. The above MHV The primers are fused to a DNA sequence that encodes a mouse Η chain leader sequence, and the above MHC primers are C chain C region: DNA sequence fusion. In addition, all 5 / terminal primers (MKV and MHV) contain a sequence GTCGAC that can provide a restriction enzyme Sa 1 I cut near the 5 &gt; end, and all of them 3 / Terminal primers (MKC and MHC) are those containing a nucleotide sequence CCCGGG that can provide a restriction enzyme Xm a I cut near the 5-terminus. These restriction enzyme cuts are made so that they can encode two V regions The target DNA fragment is used for asexual propagation in various selection vectors. If these restriction enzyme cutting sites also exist in the target DNA sequence that can encode the two V region, only It can also be used as a carrier, or it can be other restriction enzyme cutting site.

(4) Isolate the DNA that can encode the V region Secondly, in order to obtain the DMA fragment that can encode the V region of the mouse monoclonal antibody, low-melting agarose or layered column (refined pc R product kit (QUGEN, etc.) , DNA purification kit (GENECLEANII), etc.] (Please read the precautions on the back before filling this page)

This paper size is in accordance with Chinese National Standard (CNS) A4 specification (210X297 mm) _. At Γ 416960 B7. V. Description of invention (15) It is separated and refined P C R enlarged product. On the other hand, the restriction enzymes Sa 1 I and Xma I were subjected to an enzyme treatment to obtain a DNA fragment encoding a target V region of a mouse monoclonal antibody. On the other hand, by using the same restriction enzymes Sa 1 I and Xma I to cut the appropriate selection vector such as plastid P UC 19, the above-mentioned DNA fragment is enzymatically linked to pUC19 to obtain a mouse that can encode A plastid formed from a DNA fragment of the V region of interest of a monoclonal antibody. The DNA sequence after selection can be determined by any conventional method. For example, an automatic DNA sequencer (manufactured by Applied Biosystems Inc.) can be used to determine the DNA and sequence determination of the purpose of the selection in Examples 1 and 2. Complementarity-determining region (CDR) The present invention further provides a hyper-V region or complementarity-determining region (CDR) of the V region of a mouse monoclonal antibody against human IL-8. The L chain of the antibody and the V region of the Η chain form the antigen binding site. This region on the L and Η chains has a similar basic structure. The two-chain V region contains four frame structure regions that are compared for sequence comparison. These are linked by three hyper V regions or CDR. (Kabat, Ε.Α., etc., "Sequences of Proteins of Immunological Interest", US Dept. Health and Human Services 1991) 0 Most of the above four framework structural regions (FR) have Lu-lamellar structures, three The CDRs are formed as circles. Sometimes CDRs can also form part of a point-to-sheet structure. The three CDRs are kept in three-dimensionally close positions with each other by FR, and are then paired with the three CDRs—from the paper size to the Chinese National Standard (CNS) A4 specification (210X297 mm), 18 _ A7 B7 416960 V. Description of the invention (16) Play a role to form an antigen binding site. Please read the notes below 1¾ and then fill out this page. The present invention also provides these CDRs, which are extremely useful as materials for human-type antibodies, and DNAs encoding these. These CDR regions can be compared with the known amino acid sequences on the V side, and can be determined according to the empirical principles of "Sequences of Proteins of immunologicai I. uteres dagger" such as Kabat, E.A., etc. The specific description is listed in the examples. 3. Production of Chimeric Antibodies Printed by the Real Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs Before designing the anti-human IL-8 reconstituted human V region, it must be confirmed that the CDRs used can actually form antigen-binding regions. Chimeric antibodies are made here for this purpose. In order to make a chimeric antibody, it is necessary to first establish a DNA that encodes the L chain and Η chain of the chimeric antibody. The basic method for establishing the two DNAs is to link each DNA sequence of the mouse leader sequence and mouse V-region sequence that can be seen when PCR-pure germline c D Ν Α is connected to a mammalian cell expression vector that already exists human C Zone DN A sequence. The human antibody C region can be any human L chain C region and any human Η chain C region. For example, the L chain can be the human L chain Cx region or CA. When the Η chain is I gG, it can be C r 1 C? * 2, Cr3 or Cr4 (Ellison, J. et al., DNA, 1, 11-18 (1981), Takahashi, N, et al., Cell, 29, 671-679 (1 9 8 2) , Krawinke 1, U., etc., EMB0, J. 1, 4 03-407 (1982)) or other similar types. In order to make chimeric antibodies, two expression vectors must be made, that is, they can be encoded in mice under the control of expression control regions such as enhancer / promoter system. The paper size is applicable to China National Standards (CNS) A4 specifications (210 X 297 mm) -19-A7 乂 41 6960 B7_____ V. Description of the invention (l7) Expression vectors made from DNA of L chain V region and human L chain C region, and expression control in enhancer / promoter systems and the like Under the control of the region, it contains the expression vector formed by DNA encoding mouse V chain and human NA chain C. Second, these two expression vectors are used to simultaneously transform host cells, such as mammalian cells, and then culture the transformed cells in vitro or in vivo to produce chimeric antibodies (eg, W09 1-1928). Or introduce the DNA encoding the mouse L chain V region and the human L chain C region and the DNA encoding the mouse limulus chain V and human limulus chain C regions into a single expression vector, and then use the vector to transform the host cell, This transformed host is then cultured in vitro or in vitro to produce a chimeric antibody. In Example 4, it was described that a chimeric anti-skeleton was produced from the single bead antibody WS-4. The PC R method was used to clone the DNA encoding the mouse ws-4 X-type L chain leader region and VM c DNA and ligate it to a expression vector containing a human gene D NA encoding the human L chain C X region. Using the PCR method, the c DNA encoding the Η chain leader region and the V region of the mouse WS-4 antibody was also cloned and linked to a expression vector containing genomic DNA encoding the human C r-1 region. Printed by the Central Laboratories of the Ministry of Economic Affairs and the Consumer Cooperative. More specifically, using specially designed PCR primers, the cDNA encoding mouse ONS-M2 1 V ， was introduced with appropriate bases at its 5 and 3 1-termini. Sequence, (1) make it easier to insert into the expression vector, and (2) make them function properly in the expression vector (for example, in the present invention, the introduction of Kozak sequences to improve human transcription effectiveness). Secondly, the V region of mouse WS-4 obtained by PCR amplification using these primers is inserted into the paper size that already contains the desired human C region. The Chinese national standard (CNS &gt; A4 size (210X297 mm) A7 I * * 41 6960 B7 V. Description of the invention (18) HEF expression vectors (Figure 1). These vectors are very suitable for the transient or stable expression of genes whose antibodies are manipulated in various milk cell lines. Test the binding activity of the chimeric WS-4 antibody prepared as described above, and the chimeric WS-4 antibody system has the activity to bind to human IL-8, (Figure 2) Therefore it shows that the correct mouse V region has been It has been purified and its sequence has been determined. Designed to reconstitute human WS_4 antibody to make a mouse monoclonal antibody. The CDRs of the human antibody are transplanted to the human antibody. The amino acid sequence of FR is extremely similar to the amino acid sequence of the human monoclonal antibody FR of the transplanted CDR. It is printed by the consumer cooperation of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back first) (Fill in this page) To achieve this By comparing the amino acid sequence of the mouse monoclonal antibody FR with the amino acid sequence of the human monoclonal antibody FR, the human V region can be selected as the basis for the design and reconstruction of the human WS_4 antibody V region. Specifically, The gene chain analysis software GENETX (Software Development. Co., Ltd.) can be used to compare the L chain and Η chain VM of mouse WS-4 antibody with all known humans found in the basic data of National Biomedical Research Foundation (NBRF). V region. When compared with the known human antibody L chain V region, the mouse WS _ 4 antibody L chain V region is most similar to the human antibody HAU (Watanabe, this paper applies Chinese National Standard (CNS) A4 specifications ( 210X2.97 public compliance :) 21-A7 B7 416960 V. Description of the invention (19) S. et al. Hoppe-Seyler's Z. physiol, Chem. 3 5 1, 129 1 ~ 1295, 1970) The identity is about 69.2%. On the other hand, when compared with the known human antibody Η chain V region, the WS-4 antibody Η chain V region is most similar to the human antibody VDH2 6 (Buluiiela., L., etc. , EMB0 J. 7, 2 0 3 ~ 2 0 10, 1988), about 71.4% identity. Generally, for human V region The amino acid sequence of the amino acid sequence of the mouse V region is lower than that of the amino acid sequence of the mouse V region. This means that the mouse w S-4 antibody has no V region. It is completely similar to the human V region. At the same time, in order to solve the immunogenicity problem of the patient (human), it is best to make the V region of the mouse WS-4 into humanize. The V region of the mouse WS-4 antibody was defined by Kabat, EA, etc. (1 991) Sequences of Proteins of Immunological Interest. Forth Edition, US Department of Health and Human Services, U.S. Government Printing Office The common sequence of human V region subgroups is compared to FR of V region. The results are shown in Table 1. 阅读 Please read the notes on the back. I The paper printed by the Central Standards Bureau of the Ministry of Economic Affairs, Shellfish Consumer Cooperative Co., Ltd. is again applicable to the Chinese National Standard (CNS) A4 specification (210X: 297 mm)-μ A7 B7 416960 V. Description of the invention (20) Table 1 Identities between FR 'of mouse WS-4 V1S and FR of common sequences of human VMs of various subgroups (%) (A) FR in V region of L chain HSGIHSG II HSG III HSG IV 64. 4 5 1.3 5 7.3 57.5 (B) FR HSGIHSG II in H chain V region HSGIHSG II HSG III 46.9 40.9 62.3 Common sequence mouse in chain V region WS-4 Ministry of Economic Affairs of Η chain V region Subgroup of W Zone printed by the Consumer Standards Cooperative of the Central Bureau of Standards (6 4.4% FR is the most similar to human sequences, human antibody group I, and the subgroup of human chain V zone, which belongs to reconstructed human WHSG III ) Identical to known HSGI of the L chain V region S-4. Η like chain V seems to have 6 2 fruit line can support H A U antibody V group ΙΠ. Human anti-human antibodies to subgroup I (S-4 antibodies also interact with humans

L) FR of the body In addition, 另外 .3% of the L chain D Η 2 designed heavy HSG body Η chain body Η chain L chain VL chain VFR is the most similar to human L chain V +, with group III (HSG III) in common. Compared with human antibodies, the result obtained is that the V region belongs to the sub-6 of the human L chain V region, and the human V chain is derived from the human L chain V region of human WS-4 antibody. When designing the V region, it belongs to the subgroup III (V region is the best. In comparison, the mouse WS_4 antibody region is a member of the subgroup I human anti-paper standard applicable to Chinese national standards (CNS ) Α4 size (210 × 297 mm) -23-A7 [r 4 1 6960 Β7-Printed by the Consumers' Cooperative of the Central Economic and Technical Bureau of the Ministry of Economic Affairs of the People's Republic of China 5> Invention Note (21) The L key V area of the body REI is similar. Therefore, the design and reconstruction Human ws-4 antibody uses the FR of R E I in the L chain V region. According to REI, these human FRs are related to the original human EE IP a 1 τη, W and other Ho PP e-Sey 1 er, SZP hysi 〇1 Ch em ,, 356, 167- 191, 1 975; Epp, 0. etc., Bochemist η T &gt; 1 4} 4943-4952, 1975) Compared with five amino acids (position 3 9 &gt; 7 1 1 0 4 1 0 5 and 1 7 refer to Table 2) 0, The acid number of the amine-based data by 1C a b at} E. A et al. (1991) by the subject. The changes of the two amino acids in positions 3 9 and 7 1 are returned to the changes of amino acids in the FR of the L chain V ίρ5 ~ present in the mouse CAMPATH -1H anti-Bι »body (Riechm an η et al. 1 9 8 8) ° According to Ka bat et al. (199 1), the changes of the three amino acids in FR 4 (positions 1 0 4 1 0 5 and 1 7) are based on the J region of other human KL chains Still belong to humans 0 Designed to reconstruct two versions of human WS-4 antibody L chain V region &gt; FR in version RVL a and FR based on reconstructed human CAMPATH -1Η anti-M # body FR ( Ri ech ra a η η et al. 1 9 8 8) Identical 5 and CDRs are the same as CD R in L chain V of mouse WS-4 antibody 0 First * version RVL b is based on RV La Only human FR 3 One position 7 1 Amino acid is different 0 such as C. Ch 0 t hi a etc., J · Mol. Bio 1. (1 9 8 7) '1 9 6: 9 0 1 — 9 1 7 residue 7 1 defined It is the standard (CO η ο nica 1) of the CDR 1 of the L chain V region ip ff * Weighing part 0 According to the prediction, the amino acid at this position will directly affect the paper size of the L chain V zone. The Chinese national standard (CNS &gt; A4 specification (210X297 mm) ^ Please read the precautions at the back before filling «Write this page A7 I ** 41 6960 _ B7 '_ ^ _ V. Description of the invention (22) The structure of the CD R 1 coil, so it is speculated that it will greatly affect the binding of antibodies. Reconstruct human WS-4 antibody L chain V The phenylalanine at position 71 in the RV L b region was changed to tyrosine. Table 2 shows that the L chain V region of the mouse WS-A antibody was modified to be used in the reconstruction of the human CAMPATH-1H antibody. FR of RE 1 (Riechmann et al., 1898), and reconstruction of each amino acid sequence of two versions of the L chain V region of the human WS_4 antibody. Central Standards Bureau, Ministry of Economic Affairs, Shellfish Consumer Cooperatives, printed paper standard ^ Applicable in centimeters at country I and country I at C 4 1 6960 B7 V. Description of invention (23) 2 Table reconstruction human WS — 4 L chain V District Design 12 3 4 12345678901234567890123 45678901234 567890123456789

WS-4L DIQMTQSPASLSASVGBTVTITC RASBIIYSYLA WYQQKQGKSPQLLVY RBI DIQMTQSPSSLSASVGDRVTITC WYQQKPGKAPKLLIY RVLa DIQMTQSPSSLSASVGDRVTITC RASEIIYSYLA WYQQKPGKAPKLLI --------------- RVLb ------ ------------ FR1 CDR1 FR2 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5 6 7 8 9 0123456 78901234567890123456789012345678 901234567

WS-4L NAKTLAD GVSSRFSGSGSGTQFSLRISSLQPBDPCSYYC QHHFGFPRT

REI GVPSRFSGSGSGTDFTFTISSLQPEDIATYYC

RVLa NAKTLAD GVPSRFSGSGSGTDFTFTISSLQPEDIATYYC QHHFGFPRT RVLb -------- -------------- Y ----------------- ----- ---- CDR2 FR3 CDR3 10

8901234567 WS-4L FGGGTKLELK REI FGQGTKVBIK RVLa FGQGTKVEIK RVLb ---------- FR4 This paper uses China National Standard (CNS) A4 (210X297 mm) —26-| = 416960 A7 * B7 V. Invention Description (24)

Note: The F R of RE I can be found in reconstructed human CAMPATH-1H antibody (Riechmann et al., 1898). The five underlined amino acids in the FR of RE I are amino acids whose amino acid sequences are different from those of human REI. The F R line in the V region of the mouse WS-4 antibody WS chain is most similar to the human Η chain V region belonging to subgroup III (Table 1). Compared with the known human Η chain V region, the mouse WS _ 4 antibody Η chain V region is from FR 1 to FR 3, and is a human antibody VDH2 6 Η chain V region which is a member of one of the human Η chain V region subgroups ΙΠ (BuluweU, L. et al., EMBO J., 7, 2003 ~ 2010, 1988) are most similar. The FRA sequence of the FR 4 series VD Η 26 is not clear, so the human antibody 4 B 4 (San z,!. Et al., J. Immunol., 142, 883 — 887, 1989) belonging to subgroup III was used. Amino acid sequence. These human Η chain V regions were used as the basis for designing and reconstructing the human WS-4 antibody Η chain V region. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs. Designed to reconstruct eight versions of the V region of the human WS-4 antibody chain. Of the eight versions, human FR1, 2 and 3 are based on the human antibody VDH2 6, FR1, 2 and 3, and FR4 is based on the human antibody 4 3 4? The ruler 4 and the mouse ruler 00 are the same as the CDRs of the V region of the ^ 1 chain of the mouse 3-4 antibody. Tables 3 and 4 show the mouse WS_4 antibody Η chain V region, the template human antibody VDH2 6 FR1 ~ 3, human antibody 4 Β 4 FR 4 and reconstructed human WS-4 antibody Η chain V region eight Versions of each amino acid sequence. 27 This paper size applies the Chinese National Standard (CNS) A4 specification (2 [0 × 2.97 mm &gt; _ V. Description of the invention (25) * A7 B7 f 416960 Reconstruction of the design of the human WS-4H chain V zone. 1 2 3 WS-4H 123456789012345678901234567890 EVKLVESGGGLIQPGDSLRLSCVTSGFTFS 12345 DYYLS VDH26 RVHa ~h EVQLLESGGGLVQPGGSLRLSCAASGFTFS EVQLLBSGGGLVQPGGSLRLSCAASGFTFS DYYLS FR1 CDR1 central Bureau of standards, Ministry of economic Affairs J Engineering Co-op printed WS-4H VDH26 RVHa RVHb RVHc RVHd RVHe RVHf RVHg RVHh 4 67890123456789 WVRQPPGKALBWVG WVRQAQGKGLELVG WVRQAQGKGLBLVG ------ ---------- p -------- ----- P ----- W-- ____ pp _____— ——- P—A--W ------ ------------ W-- FR2

5 6 012ABC3456789012345 LIRNKANGYTREYSASVKG LIRNKANGYTREYSASVKG CDR2 This paper size applies to Chinese National Standard (CNS) A4 specification (210X297 mm) 00 &quot; ίο 5. Description of the invention (26) Table A7 B7 (ί: 41 6960 WS-4H VDH26 RVHa RVHb RVH RVHe RVHf RVHg RVHh

7.8 9 67890123456789012ABC345678901234 RFTISRDDSQSILYLQMNTLRGEDSATYYCAR RLTISREDSKNTLYLQMSSLKTBDLAVYYCAR RLTISRBDSKNTLYLQMSSLKTBDLAVYYCAR

100 567890ABC12 ENYRYDVELAY ENYRYDVELAY Please read the notes at the back

i WS-4H 4B4 RVHa ～ h

-F ---------- 110 34567890123 WGQGTLVTVSA WGQGTLVTVSS WGQGTLVTVSS FR3 CDR3 Note: RVHa ~ h represents RVHa, RVHb, RVHc, RVHd, RVHe, RVHf, RVHg, and RVHh Central Laboratories Consumers Cooperatives, Ministry of Economic Affairs Printed on paper

R P i mm 7 9. 2 Printed by the Consumer Cooperatives of the Central Procurement Bureau of the Ministry of Economic Affairs Α7 Γ 41 6960 _ B7 V. Description of Invention (27)

DN A, which encodes the reconstituted human WS-4 antibody V region, was specifically described in Example 5. V, which synthesizes and encodes the reconstituted human WS-4 antibody L region and V chain, each The first version of DNA. The sequence was then determined to confirm that the entire DNA sequence of the reconstructed human WS-4 antibody L chain and Η chain V region version "a" could encode the correct amino acid sequence. The sequence of the reconstructed human WS-4 antibody L chain V region version "a" is shown in Sequence 6 2 and the sequence of the reconstructed human WS-4 antibody L chain V region version ^ a "is shown in Sequence 38. Other versions of DNA that can encode the reconstituted human WS-4 antibody V region use the first version "a" as a template and use a slightly modified published PCR-mutation induction method (Kamman, Μ et al., Nucleic Acids Res · ,, 17, 5 4 0 4, 1 9 8 9). As described in the design of the reconstruction of the human WS-4 antibody V region, a DNA encoding an additional version (version "b") of the reconstructed human WS-4 antibody L chain V region was produced, and then there was an encoding of the reconstruction human WS-4. Seven additional versions of the antibody V-chain V region (versions "b", "c", "d", "ej, rf", "g", and "hj)". These additional versions contain information from the first In the version, there is a series of small changes in the amino acid sequence. These fine changes in the amino acid sequence can be achieved by applying PC R mutation induction method to the DNA sequence. Introduce the necessary PCR primers that change in the DNA sequence. After a series of PCR reactions, the PCR product is then germlined, and then the sequence is determined to confirm the changes in the DNA sequence. The paper scale is applicable to the Chinese national standard (匚 肥) 6 4 Specifications (210 father 297 mm) _3〇 (Please read the notes on the back before filling in this card)

at f &quot; 41 6960 B7 ’5. The invention description (28) has been produced as planned. The sequence of the reconstructed human WS-4 antibody L chain V region "bj" is shown in sequence 65, and the reconstructed human WS-4 antibody Η chain V region versions "b", "c", "d", "e", "f The sequences of "", "g", and "h" are shown in the sequences 4 1, 4 4, 4 5, 48 »51, 54» 550, and the sequence determination of the DNA sequence of various versions of the human WS-4 antibody VE is confirmed. Then, the DNA encoding the V region of the reconstituted human WS-4 antibody was subcultured into a milk cell expression vector that already contained the DNA encoding the human C region. That is, the DMA encoding the reconstructed human WS-4 antibody L chain V region is linked to the DNA sequence encoding the human L chain C region, and the DNA encoding the human WS-4 antibody Η chain V region is linked to the human r-1 1 DNA sequence of C region. Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs (please read the notes on the back of the page before filling out this page). Next, test the reconstruction of the human chain L region V version "a" or "'b". The combination of all the combinations "~ h" on human IL-8. The results are shown in FIG. 7, showing that two types of reconstructed human antibodies (RVLa / RVHg and RVLb / RVHg) containing L chain versions “a” and “b” and 」chain version“ h ”have chimeric WS — 4 The antibodies bind to IL-8 activity at the same level. In order to produce the chimeric antibody of the present invention against human IL-8 or reconstituted human antibody, any expression line can be used, such as sclerotinia cells, such as animal cells, such as established saccharomyces militaris cell line, spore cells, and yeast cells. , And prokaryotic cells, such as bacterial cells, such as coliform cells. Preferably, the chimeric antibody or reconstituted human anti-system of the present invention is expressed in mammalian cells, such as COS cells or CHO cells. This paper size applies to the Chinese National Standard (CNS) A4 specification (2) 0 X 297 mm _ w _ ^ 416960 A7 B7 Printed by the Consumer Cooperatives of the Central Bureau of Standards, Ministry of Economic Affairs ^ 5. Description of the invention (29) In these cases The following J can be used for mammalian cell performance. Commonly useful promoters can be used. For example, the use of human cell megavirus prophase (hu ina η cy t 〇nt ega 1 cvinusiu ni ediateear 1 y; HCMV). Examples of promoter expression vectors include: CM V — V Η — HC r 1 9 Η CM V — VL Η CK and other sources including PSV 2 η e 0 (see International Publication W 0 9 2 — 1 9 7 5 9) 0 Furthermore, other promoters of gene expression in mammalian cells that can be used in the present invention can also use viral promoters such as retrovirus polyoma virus adenovirus simian virus 4 0 (CV 4 0). Or human • peptide «bond • extension · factor — 1 a (Η ET — 1 a), such as the promoter 0 of white milk mammal cells, for example, when using the promoter of SV 4 0, the method of Mu 1 1 iga η &gt; R. C. etc. (Na t U re 2 7 7 J 1 0 8 1 1 4, 1 9 7 9), and also use Η EF — 1 a promoter can follow the method of Mizushima) S. (Nu cl ei C Ac id S Re S.) 1 8 5 3 2 2 1 9 9) 0 Other specific examples of promoters useful in the present invention are the EF-1a promoter. The expression vector containing this promoter contains Η EF — V Η — gr 1 and Η EF-VL — g K (Figure 1). 〇 Copy from EGJ J-, eggplant can be used to white polyoma virus, adenovirus , SV 4 0 ί The DNA sequence of bovine papilloma virus (BP V) can be used to amplify genes in host cell lines by 1 y, and use amino grapes as selection targets. -F— «· 甘3 〆—Phosphotransferase or η e 〇rrr 1. Tolerance gene Thymic nucleus-t-μGlycokinase (TK) gene, coliform xanthine-guanine to r VJ phosphoribosylase (XGPRT) Please read τ6 first Note S (Writing on this page The paper size applies to the Chinese National Standard (CNS) A4 size (210 X 297 mm) _ 3 2 Bu A 1 6960! Printed by Guigong Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs (30) Gene two gasification folate reductase (dhfr) gene 0 In summary 5 The present invention provides first Human IL-8 mouse monoclonal antibody L chain V region and Η chain V region, and DNA encoding the L 'chain V region and DNA encoding the Η chain V 0 These are for the production of anti-human IL — Human / mouse chimeric antibodies and reconstructed human antibodies of 8 are extremely useful. 0 As a single anti-rtrrffa body, it can be a WS-4 0 L chain V region having the amino acid sequence J shown in sequence 2 6 and Η The chain V region has, for example, the amino acid sequence 0 shown in the sequence 2 7 These amino acid sequences are respectively encoded by the nuclear +-* · acid sequence shown in the sequence 2 7 2 7 The anti-human IL of the present invention — The chimeric antibody system of 8 is called (1) human L chain C region and small wind L chain V region and (2) human 2 chain C region and mouse Η chain V region are called 0 mouse L chain V region and small The V-region of the murine chain and the DNA encoding them are as described above. The above-mentioned human L-chain C The region may be any human L chain C region. The above-mentioned human Η bond C region may be any human Η chain C region. It may be, for example, CT 1 &gt; C r 2, C T 3 or CT 4 (E 1 1 i SO η, J, etc., DNA, 1 1 1 — 1 8 (1 9 8 1) 9 Ta ka ha shi, N *, etc. Ce 11, 2 9 &gt; 6 7 1 — 6 7 9 (1 9 8 2) 9 Kr aw i τιke 1, U. Et EBM0 J. 1, 4 0 3 — 4 0 7 (1 9 8 2)) Region 0 Two expression vectors are produced for the production of chimeric antibodies, namely by promoter / promoter system Under the control of a performance control surface, a performance vector made of DNA encoding human L chain is produced, and the paper size of the promoter / promoter system is applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm) _ 33 A7 B7 ί 416960 V. Description of the invention (31) The expression vector made of DNA that can encode the human chain is produced under the control of the expression control area of (31). Secondly, by using these expression vectors to simultaneously transform host cells such as milk cells, the transformed cells are cultured in vivo or in vitro to produce chimeric antibodies. It is also possible to introduce DNA encoding mouse L chain V blue and human L chain C region and DNA encoding mouse Η chain V region and human Η chain C region into a single expression vector, and use the vector to transform host cells Then, this transformed host is cultured in vivo or in vitro to produce a chimeric antibody. The reconstituted WS-4 antibody system of the present invention consists of (A) an L chain containing (1) a human L chain C region, and (2) a human monoclonal antibody L chain CDR containing a human L chain FR and an anti-human IL-8 L chain formed by the V region; and (B) a V chain containing the V chain of the human Η chain C and (2) a 单 chain CDR containing the human Η chain FR and a human monoclonal antibody against human IL-8 Made into a chain; made into. A preferred embodiment printed by the Central Bureau of Standards, Ministry of Economic Affairs, Consumer Cooperatives is the amino acid sequence shown in the above L chain CDR sequence 26. The amino acid sequence has the amino acid sequence defined in Table 5, The fluorene chain CDR has an amino acid sequence shown in sequence 27, and the amino acid sequence ranges from the amino acid sequence defined in Table 5; the human L chain FR is from a human being; the human η Chains FR1, 2 and 3 are from VDH2 6, FR4 are from 4B4; the above human L chain C region is human c X region; and the above human chain C region is human. The paper standard is applicable to Chinese National Standard (CNS) A4 specifications (210 × 297 mm &gt; _ Γ 416960 Α7 ^ '___ Β7 ______ V. Description of the invention (32) C r 1 region. Instead of the above-mentioned human L chain V region and / or human thorium chain V region, radioisotopes can also be combined. For reconstituted human antibodies with sufficient activity for specific antigens, it is best to replace a part of the amino acid sequence of the human FR described above. The Η chain V region has RVHa shown in Tables 3 and 4. , RVHb, RVHc, RVHd, RVHe, RVHf, RVHg or RVHh amino acid sequence. In addition, the amino acid at position 41 in the FR2 of the H chain is proline, and the amino acid at position 4 to 7 is tryptophan. And / or the amino acid at position 6 or 7 in the FR3 is preferably phenylalanine, and it is more preferable to have the amino acid sequence shown by RVHb, RVHd, RVHe, RVHf, RVHg or RVHh, among which RVHg is most suitable as H chain V zone. Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs to produce two expression vectors for the production of reconstruction antibodies, that is, under the control of expression control zones such as promoters / promoters, production of coded reconstruction human L The expression vector formed by the DNA of the strand, and the expression vector composed of D Ν Α which can encode the reconstituted human Η chain under the control of the expression control region such as the promoter / promoter system. Host cells transformed into milk-like cells, cultured in vitro or in vitro to produce reconstituted human antibodies. DNA encoding the reconstituted human L chain and DNA encoding the reconstituted human chain can be introduced into a single expression Vector, use this vector to transform the sink The main cells are then cultured in vitro or in vivo to produce reconstituted human antibodies. 35 This paper is in accordance with the Chinese National Standard (CNS) 8.4 (2 [〇 × 2.97 mm) Α7 Β7 Central Ministry of Economic Affairs Printed by the Employees' Cooperatives of the Hong Kong Standard Bureau. 416960 V. Description of the invention (33) Chimeric antibodies or reconstructed human antibodies produced as described above can be used according to conventional methods, such as protein A affinity chromatography, ion exchange chromatography, gel Refinement by filtration and other methods. The chimeric L chain or reconstituted human L chain of the present invention can be combined with a limulus chain to be used to make a more complete antibody. Similarly, the chimeric bond or reconstituted human chain of the present invention can be used in combination with the L chain to make a more complete antibody. The mouse L-chain V region, the reconstructed human L-chain V region, the mouse limulus-chain V region, and the reconstructed human limulus-chain V region are regions that can originally bind to the antigen human IL-8. In itself, or as a fusion protein formed with other proteins, it is extremely useful for medicine and diagnostic medicine. In addition, the CD chain of the L chain V region and the CDR of the limulus chain V region of the present invention are originally a part capable of binding to human IL-8 of the antigen, and may be a fusion protein by itself or with other proteins, which are used as medicines and diagnostic drugs. Extremely useful. The DNA that can encode the mouse L chain V region of the present invention is extremely useful in making DNA that encodes the embedded L chain, or DNA that can reconstruct the human L chain. Also, DNA, which can encode the V region of the mouse cymbal chain of the present invention, is extremely useful for making DNA that can encode chimeric cymbal chains, or DNA that can reconstitute human cymbal chains. In addition, the DNA encoding the CDR of the L chain V region of the present invention is extremely useful in making DNA that encodes the reconstruction of the human L chain V region, or DNA encoding the reconstruction of the human L chain. In addition, a suitable host can be used to generate a human antibody F (ab &lt;) 2, Fab or Fv, or a single chain F ν of two types F ν of Η chain and L chain can also be used for the above purpose (for example, refer to this Paper size applies Chinese National Standard (CNS) Α4 specification (210X297 mm) (Please read the notes on the back before filling this page)

36-V. Description of the invention (34) A7 B7 f, 4 1 6960, Bird, RE, etc., TI.BTECH, 9, 1 3 2 — 1 19 9 1) 0 Single chain F v V region and L chain V are borrowed Single-chain F is formed by linking the anti-human I region with a linker, preferably through peptides S. et al., Pr Oc. Natl, Acad. Sc 5879-5883, 198 single chain F v Region and any reconstituted human antibody L chain and L chain 38,41,44 &gt; 45,4 The L chain formed by the amino acid sequence described in one of these V regions is preferably linked by a peptide such as amino acid 1 2 ~ 1 The DNA of the single-stranded F ν encoded by residue 9 is the DNA of the Η chain of the human antibody or the V region of the Η chain V region as a template. For the acid sequence DNA, the PCR method is used to amplify the DNA. D N Α and each chain at both ends &lt; &gt; Once the expression vector containing the expression vector and the host can be prepared by encoding a single method, the host can be used to reconstitute the human antibody Η chain ν in the single-chain F ν system and the antibody molecule phase L-8, Η The chain V area is connected to the L chain V area connector (Hustqn, J.i.U.S.A., 85, 谙 Read the notes on the back and write this printed by the Consumer Cooperatives of the Central Procurement Bureau of the Ministry of Economic Affairs 8) 0 L chain VV region 0 8 and 5 V region are connected to each other so as to be controllable at both ends by D N A codeable, L chain region may be 1. 5 as single specific F Knot. Peptide-linked single-stranded peptide. Encoding the above, and encoding the above is the containing sequence. Any of the 5 ν ° adaptors can use the described reconstruction of the L chain or L chain of the desired amino pair primers in the sequence, made by the peptide linker unit The DNA encoding the linked pair of primers to the strand Fv can be used to obtain the single-stranded F ν by the usual method transformed by these expression vectors. Better than the activity of migrating to the organization. This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm). 37 Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs printed m J 416960 A7 B7. 5. Description of the invention (35), you can It is expected to be used for imaging made by radioisotopes and as a therapeutic agent having the function of rebuilding human antibodies. In order to confirm the chimeric antibody of the present invention, and reconstitute the binding activity of human typed antibodies and their F ab, F v or single chain F v to human IL-8, EL I SA (enzyme-linked immunosorbent assay), e IA (enzyme immunoassay), RIA (radioimmunoassay) or fluorescent antibody method. For example, using an enzyme immunoassay to test chimeric antibodies. When rebuilding human antibodies, you can add human IL-8 to a dish containing a variety of anti-human IL-8 antibodies, and then add a chimera that produces anti-human IL-8. Antibodies, reconstituted human typed antibodies or F (abj) 2, Fab, Fv or single-chain Fv cells culture supernatants or purified samples, with appropriate secondary antibodies labeled with enzymes such as alkaline phosphatase . After the culture plate is cultured and washed, an enzyme substrate such as p-nitrophenyl phosphate is added, and the absorbance is measured to evaluate the antigen-binding activity. The anti-human IL-8 chimeric antibody, reconstructed human typed antibody and its F (ab &gt;) 2, Fab, Fv or single chain Fv on the IL-8 binding inhibitory activity of the IL-8 receptor are often borrowed. Assessed by ligand cell receptor knot inhibition analysis. For example, IL-8 binding inhibition on IL-8 cell receptors on neutrophils is separated by centrifugation or other means from neutrophils obtained from heparin blood collection, etc., and then adjusted to the number of cells suitable for the above analysis. Cell suspension was used. IL-8, appropriately labeled with 1251, etc., is dissolved with unlabeled IL-8, and a solution of the antibody of the present invention or a fragment thereof prepared at an appropriate concentration, and the above neutrophils are added thereto. Chinese standard (CNS) A4 specification (21〇 × 297 mm) _ 38 (Please read the notes on the back first and write this page) after ordering a certain amount of A $! Order A7! 4 16960 A 7. ___ B7 V. Description of the invention After (36) time, the neutrophil can be separated and the activity marked on the neutrophil can be measured. To evaluate the activity of the antibody or fragment thereof of the present invention for inhibiting neutrophil migration (Chemotaxis), Use generally known methods, such as those described in J. Biol, Chem., 2 65, 8 3 1 1 to 8 3 1 6, 1 990. When using a commercially available walking action chamber, use an appropriate method. After the antibody or fragment thereof of the present invention is diluted in the culture solution, IL-8 is added and injected into the chamber respectively. Then, the prepared neutrophil suspension is added to the chamber and left for a certain period of time. It will adhere to the filter installed in the room, and the neutrophil can be measured by ordinary methods such as staining solution or fluorescent antibody. It can also be determined by naked eyes under a microscope or automatically measured using machinery. The anti-human IL of the present invention -8 chimeric antibodies Reconstructed human-type antibodies and their F (ab &lt;) 2, Fab, Fv or single chain Fv are printed by rlv through the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page) After the filter is sterilized by filtration, it is preferably administered orally, such as intravenous injection, intramuscular injection, intraperitoneal injection, subcutaneous injection, etc., as a pharmaceutical therapeutic agent. The amount administered to a person is due to the patient's disease Symptoms, ages, etc. are different, about 1 to 100 mg per person, and a separate dosage of 1 to 10 mg per kg per week can be selected. The chimeric antibody against human IL-8 of the present invention reconstructs the human type Antibodies and their F (ab &lt;) 2, Fab, Fv, or single-chain Fv systems are purified and evaluated for binding activity. The preparations are generally used as pharmaceutical therapeutic agents by the method of preparing physiologically active substances. For example, injection The preparation is a purified chimeric antibody against human IL-8, and reconstructed human type antibody. The paper size is applicable to the Chinese National Standard (CNS) A4 specification (210X2.97 mm) _ _ A7 B7 Γ 416960 V. Description of the invention (37) (Please read the note on the back first Xiang rewrite this page) and its F ab, F v or single-chain F v are dissolved in solvents, such as physiological saline, buffers, etc., and anti-adsorbents such as Tw ee η 8 0, gelatin, human Serum albumin (HSA), etc. can also be dissolved and reconstituted lyophilized before use. For excipients during lyophilization, sugar alcohols or sugars such as mannitol, glucose, etc. can be used. The examples specifically illustrate the present invention, but the present invention is not limited thereto.

Example 1. Breeding D N A that encodes a mouse monoclonal antibody against human IL-8

Select D N A that can encode a mouse monoclonal antibody against human IL-8 as follows

1. Modulation of whole RNA Modification Chirgwin, J. M. et al. Biochemistry,! · 8, 5 2 9 4 — 5 2 9 9, 1 9 7 9 Chlorine planer density gradient centrifugation self-fusion tumor WS — 4 Modulate whole RNA. Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs, that is, to completely homogenize 1 × 107 fusion tumor WS_4 cells in 25mj? 4M guanidinium cyanate (manufactured by Fulka), and stack the homogenate in 5. On a layer of 7M chlorinated planer solution, secondly on a Beckman SW4 0 rotor, centrifuge at 3 1 0 0 r pm at 20 ° C for 14 hours to precipitate the RNA. The RNA pellet was washed with 80% ethanol, and then dissolved in 20 μM Tris-HCi? (PH 7.5%) containing 10 m M EDTA and 0.5% N-laurylsarcosinate. 5 This paper size applies the Chinese National Standard (CNS) A4 scale (210X297 mm) Α7 Γ 416960

I B7 Printed by the Central Laboratories of the Ministry of Economic Affairs, Masonry Consumer Cooperatives 5. In the invention description (38)) &gt; Then add 0 5 mg / mi? Protease (B 〇ehri ng er) and keep in a 37 ° C warm bath Constant temperature 30 minutes 0 Extraction of the mixture with phenol and chloroform 9 Precipitation of RNA with ethanol Q Second re-dissolution of the R Ν A precipitate in 2 0 0 2 (τη Μ T r 1 S Η cn (P Η 7-5) in 0 2 • The messenger R Ν A (m R Ν A) 0 was extracted to extract the mRNA encoding the white mouse monoclonal antibody WS — 4 Η chain 9 using Fas t Track mRNA I so 1 at ί ο η K it Ve rsion 3. 2 (In vitr 0g en) »According to the method described in the instruction book 9 White RNA extraction from the above 1 to τη RNA 0 3 synthesis sheet of P ο 1 y (A) CDNA is recorded in c D Ν A Cy cl e K it (manufactured by I η ν it r 〇gen) according to the instruction Method: Synthesize single-stranded CDNA from about 40 η gm RNA obtained from the above 2 and use it to amplify the cDNA encoding mouse Η chain V IE. 0 &gt; To amplify the CDNA that encodes the mouse L chain V region. 9 White approximately 1 0 g of the above-mentioned full RNA to synthesize single-stranded CDNA 〇 4 Amplify the gene encoding the variable region of the antibody by PCR (1) Amplify the sequence of the CDNAPCR method that can encode the V region of the mouse Η chain. Sequences used: 1 3 ~ 2 Primer shown in MHV (Mo US e He a ν y Va riab 1 e) shown in 4 — 1 ~ 1 2 Sequence: 2 5 Primer Μ Η C (Mo US € He a ν y Co ns tant) primer (Jo nes, s • Τ. etc. j B io / Te ch no Ιο gy 9 9 ϊ 8 8 — 8 9 This paper size applies to China National Standard (CNS) A4 (210X297 mm μ-printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs) Α7 Γ 41 6960, ____B7 '. V. Description of the invention (39) 1991. 100 &quot; The PCR solution contains 10mM Tris-HCJ? (PH8.3.), 50mM Potassium, 0.1 m M dNTPs (dATP, dGTP, dCTP, dTTP), 1.5 mM magnesium dichloride, 0.001% (W / V) gelatin, 5 units of DNA polymerase AmpliTaq (Perkin Elmer Cetus) 0.25 sequence: one of the MHV primers shown in 1 3 to 24 and 1.7 5 yM sequence: the MHC primer shown in 25 and the result obtained in 3. above. 1. Prepare MHV1 ~ 12 primers for single-stranded cDNA. After covering with 5 0 # mineral oil, the initial temperature is 94 ° C for 3 minutes, followed by 94 ° C for 1 minute, then 55 ° C for 1 minute, and 72 ° C for 1 minute. After this temperature cycle was repeated 30 times, the reaction mixture was kept at 72 ° C for 10 minutes. (2) The primers for PCR amplification of the c DNA PCR that can encode the V region of the mouse Η chain use the primers 1 ~ 1 1 of M KV (Mouse Kappa Variable) shown in sequences 1 to 1 and MKC shown in sequence 1 2 (Mouse Kappa Constant) Primer (Jones, S. T, et al., Bio / Technology, 88-89, 1991) 0 Amplified cDNA line except using 0.25vM of each MKV primer and 3.0 # M of MKC-Gl primer ΟμίThe single-stranded cDNA solution obtained in the above 3 was amplified in the same way as described in the amplification of the Η chain V region gene in 4 (1) above except for the mixture. 5. Refined PCR products and fragmentation This paper uses Chinese National Standard (CNS) A4 specifications (210 '乂 2.97 mm) _ u _ (Please read the precautions on the back before filling this page)

A7 B7 t 416960 V. Description of the invention (40) The DN A fragments of the Η chain V region and the L chain V region amplified by the PCR method as described above were separated by electrophoresis with 1.5% low melting agarose (manufactured by Sigma Corporation). . Separate a liposugar piece containing about 4 5 Q b ρ long DNA fragment and about 400 bp L-chain DNA fragment, melt at 65 ° C for 5 minutes, and add the same volume containing 2 mM EDTA and

20 OmM sodium chloride 2 OmM Tris-hydrochloric acid (pH 7.5). This mixture was extracted by phenol and chloroform, and the DNA fragment was recovered by ethanol precipitation, and then dissolved in 10 m M Tris s-HCl (ρΗ7.5) containing ImM EDTA. Secondly, 5 units of the restriction enzyme Xma I was used in 1 Tris-hydrochloric acid (pH 7.9) containing 10 μm of magnesium chloride and ImM bisthreitol.

(New England BioLabs) digestion at 37 ° C for 3 hours. Next, it was digested with 40 units of restriction enzyme Sa 1 I (manufactured by Takara Shuzo Co., Ltd.) at 37 ° C for 2 hours, and separated by agar bran electrophoresis using 1.5% low melting agarose (manufactured by S 丨 gma Co., Ltd.). Generated DNA fragments. Cut out agarose tablets containing DNA fragments, and then melt at 65 ° C for 5 minutes. Add the same volume containing 2mM EDTA and. Printed by 2 OmM Tr2, 20 OmM Affluent Sodium. is — hydrochloric acid (pH 7.5). This mixture was extracted by phenol and chloroform, and the DNA fragment was recovered by ethanol precipitation, and then dissolved in 10 m M Tris-hydrochloric acid (ρΗ7.5) containing ImM EDTA. A DNA fragment containing a gene encoding a mouse kappa L chain V region and a DNA fragment containing a gene encoding a mouse kappa L chain V region were obtained as described above. The above DNA fragments have the paper size at the 5 / -end of the paper. Applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm) _ π _ Γ 41β960ι B7 Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. ) S a 1 I cohesive end> and X ma I cohesive end at its 3 end ο 6. Concatenation and transformation make about 0-3 μ g as described above containing a codeable mouse κ-type L The Sa 1 I -X ma IDNA fragment made by the genes in the V region of the chain at 16 ° C contains approximately 0 1 μg of sa 1 IX ma I and alkaline phosphatase of E. coli (BA P Takara Sho Reaction) The PUC 1 9 vector prepared by digestion and 1 unit of T 4 DNA ligase (manufactured by GI BC0 BRL Co., Ltd.) and the attached buffer are reacted for 4 hours to link them. Next, 5 U is added. 9. Above Linking the mixture to 50 0 E. coli D Η 5 α sensor cells (GIB C0 BRL company) 7 Then let the cells stand on ice for 30 minutes &gt; at 4 2 ° C for 1 minute and then stand on ice for 1 minute 0 and then add 4 0 0 μ 9. 2 XY T medium (Mo 1 -e CU 1 ar C 1 0 Π in g: A Labor at or y Ma nua 1 1 Sa mbr 0 ok etc. Co Id S pr in g Har rb or L ab 〇r at or y Pr es S, 1 9 8 9 ) 9 to 3 7. . After 1 hour incubation, y is contained in 50 μg / m 9. amine penicillin 2 XY τ agar medium (Mo) 0 cu la Γ c 1 on in g: A La bo rat ο ry Μ an ua 1, S am br 0 0 k et ί Co 1 d S pr in g Har rb or Lab or at or y Pr ess (1 9 8 9)) seeded this coliform and cultured at 3 7 ° c— * 得 得 coliform transformation mj〇i体 0 At this time, as a selection marker, 5 0 JW g X — G a 1 (5 — bromo — 4 — chloro — 3 — fluorenyl — β — D — galactose · + · * · Gan, made by Baojiu Brewery Co., Ltd. ο The paper size is applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm) A7 B7 Γ 416960 V. Description of the invention (42) Please read the precautions on the back before filling in this% at 37 ° C. Cultivate this transfectant overnight in 10 m J2 2XYT medium containing 5 0 &quot; g / mj? Aminoamyl penicillin. Use Q1AGEN Plasmid mini-kit (manufactured by QIAGEN) according to the method described in the instruction. This culture modulates plastid DNA. The thus obtained plastid containing the gene encoding the mouse x-type L chain V region from the fusion tumor WS-4 was named PUC-WS4-VL. Except that the coliform-sensing cells were changed to JM1 0 9, they were all made by the same method from S a 1 I — Xma I DNA fragments containing genes encoding V region of mouse Η chain from fusion tumor WS-4. The plastid was named pUC-WS. Example 2 Determine the DNA base sequence using an automatic DN A sequencer (manufactured by Applied Biosystem Inc) and T aq Dye D eoxy Terminator Cycle Sequencing Kit (manufactured by Applied Biosystem Inc) according to the protocol specified by the manufacturer ^ Determine the base sequence of the c DNA codon in the above plastid. Printed by the Central Consumer Bureau of the Ministry of Economic Affairs, Consumer Cooperatives. The sequence 2 6 indicates the base sequence of the L chain VM of the mouse WS-4 antibody contained in the pUC-WS 4-VL plastid. The sequence of 7 indicates the base sequence of the gene encoding the Η chain V region of the mouse WS-4 antibody contained in plastid PUC_WS 4-VH.

Example 3 The basic structure of the CDR L bond and the V region of the Η chain is determined to be similar to each other. Each of the four frame structural parts is divided into three hypervariable regions, namely, the complementarity determining region (this paper is applicable again) China National Standard (CNS) Α4 specification (210 × 2.97 mm) Γ 416980 Α7 'Β7 _____ 5. The description of the invention (43) CDR). The amino acid sequence of the framework is better preserved, but on the other hand, the amino acid sequence of the CD R region has a high variability (Ka Sequence, EA, etc. “Sequences of P rotei π sof I mmu η ο 1 〇gica 1 I nterest "ϋ SD ept .. H ea 1 thand Human Services, 198 3) 0. According to these matters, the basic data of the amino acid sequence of the antibody made with Kabat et al. The amino acid sequence of the variable region of the mouse monoclonal antibody against human IL-8 was compared to investigate the identity, thereby determining the CD R region as shown in Table 5. 0 _5_ plastid sequence number CDR1 CDR2 CDR 3 pUC -¥ S4-VL 26 24-34 50-56 89-97 pUC-WS4-VH 27 31-35 50-68 101-111 Example 4 Confirmation of the performance of purified DNA c (Effective for making chimeric WS-4 antibody Printed by the Consumer Standards Cooperative of the Ministry of Standards of the People's Republic of China to produce a vector expressing chimeric WS-4 antibody, modified by pc R method to encode mouse WS-4 4 λ: L-chain and Η-chain VM cDNA pure pUC- WS — 4 — VL and pUC— WS 4-V Η. Then it is introduced into the HE F expression vector (refer to the above WO 92-1975). 9) (see Figure 1). The L-chain V-region post-plastid (sequence 2 8) and the chain-chain-only plastid (sequence 2 9) are fused to the first v-region leader that can be encoded. This paper size is in accordance with Chinese National Standard (CNS) A4 specifications ^ 2 丨 ϋ.97 Public love 1 ~~ -------- μ I '· 4169 60 ____B7 &gt; __ 5. Description of the invention (44) DN A, and is designed to have Kozak consensus sequences (Kozak, M, etc.) J. Mol. Biol. 196, 947-950, 1987) and Hind III restriction sites. The L-chain V-domain pre-plastids (sequence 30) and the Η-chain V-domain pre-plastids (sequence 31) are fused to a DNA sequence encoding the end of the J region, and are designed to have a conjugating donor Sequence and Bam HI restricted site. Cover with 5 0 # and mineral oil containing 2 OmM Tr is — a C n (p Η 8.3), 1 OmM potassium chloride, 6mM ammonium sulfate, 1% Triton Xl 0 0 1 0 0 ^ M dNTPs, 1. 5 m M magnesium chloride, each plastid of 1000 pm,

100 ng of template DNA (pUC-VL or pUC ^ VH) and 2.5 &quot; AmpliTaq enzyme 100mJ? PCR reaction mixture, after the first three minutes of modification at 94 ° C, at 9 4 9C for 1 minute , 30 cycles at 55 ° C for 1 minute, 72 ° C for 1 minute, and finally cultured at 7 2 ° C for 10 minutes. PC R product was purified using 1.5% low melting point agar gel and printed by the Male Workers Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs (Please read the precautions on the back before filling this page}

Hind III and BamHI were digested, and then the L chain V region was cloned in the HEF expression vector HEF-VL-gx, and the H chain V region was cloned in the HEF expression vector HEF-VH-gr. After determining the DNA sequence, the plastids containing the DNA fragments containing the correct DNA sequence were named HEF— chWS4L — g «&gt; HEF — chWS4H — gr 1 0 The infection was transferred to COS. The Chinese paper standard (CNS) A4 specification (210X297 mm) 47 Γ '416aa〇Α7 _'_ Β7 ___ 5. Description of the invention (45)

To observe the transient performance of the chimeric WS-4 antibody, the expression vector was tested in cos cells. HEF-c hWS 4 L-g λ: and H.EF-c hWS 4 L-g r L Gene Pulser (manufactured by BioRad) were simultaneously transformed into COS cells by the _ e 1 ectroporat i on method. 10 vg of each DNA was added to PBS at 1 x 107 cells / mj? In 0.8 m and divided into sections, and pulses were applied at a capacity of 1.5 x V, 25 #F. After a recovery period of 10 minutes at room temperature, the cells to which electroporation was applied were suspended in a 15 mJ? DMEM culture solution (manufactured by GIBC0) containing 5% bovine fetal serum without r-globulin. After 96 hours of incubation, the culture supernatant was collected, and the cell debris was removed by centrifugation. A disc-shaped filter with a diameter of 0.4 5 # 1 «(

Filtered by Gelman Science.

ELISA prepares EL IS A plates for measuring antigen binding and measuring antibody concentration as follows. The E L 1 S A disk for testing antigen-binding activity was prepared as follows. S goat anti-human IL was dissolved in 96-well culture plates at a concentration of 2a g / mi solidification buffer (0.1 M sodium bicarbonate, Q. 0.02% sodium azide). — 8 strains of antibodies (manufactured by r & d Systems) were solidified, and diluted with 200 &quot; 芡 dilution buffer (50 mM Tris-HC 芡 'PH7.2, 1% bovine serum protein (BSA), 1 mM magnesium dichloride, 0.15M sodium chloride, 0.05% Tween 2 〇 '0.02% sodium azide) and blocking (blocking), this paper uses China National Standards (CNS) A4 specifications (2 丨 0X297 mm) 48

416960 V. Description of the invention (46) Add 100 Renyi concentration of 5ng / mJ? Recombinant human IL · 8 (manufactured by Araersham). Sequentially dilute the purified sample of the chimeric antibody, or the cultured C 0 S cells that have been expressed. The clear solution is 'added to each well', followed by the addition of 100 M J2 concentration 1 # g / mj? Of alkaline phosphate Esterase-labeled goat anti-human IL-8 antibody (manufactured by TAGO) was' cultured and washed, and then a matrix solution (1'-nitrophenylphosphonic acid) was added. Then the absorbance at 405 nm was measured. The concentration of the antibody was determined by laminating 9 6-well culture plates with 100 # concentration 1 # goat anti-human I gG antibody (manufactured by TAGO). After blocking, a purified sample of the chimeric antibody was sequentially used. Or, make the above-mentioned C ◦ S cell culture supernatant, add to each well, and then add 1 0 0 # alkaline concentration of 1 M g / mi2 alkaline phosphatase labeled goat anti-human IL-8 antibody ( (Manufactured by TAGO), after culturing and washing, a matrix solution (1 n / mj ?, p-nitrophenylphosphoric acid) was added, and the absorbance at 40 nm was measured next. Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. Since the chimeric antibody WS_4 can specifically bind to IL-8, it can be seen that this chimeric antibody has the correct structure of the mouse monoclonal antibody WS-4V region (see Figure 2). ). In addition, the above-mentioned coliforms with plastid Η EF — c.hWS 4 L — g / c are Escherichia coli DH5a (HEF — chWS4L — g κ), and the above-mentioned coliforms with the above-mentioned plastid HEF_chWS4H — gr! Are E. coli JM109 (HEF- chWS4H-gri), in accordance with the Budapest Treaty, the Chinese National Standard (CNS) A4 is applied at 199 paper standards (2 [0X297 mm) 49

f '4 1 6960 V. Description of the invention (47) (Please read the precautions on the back before filling in this page) July 12, 4th, FERM BP — 4739 and FERM BP — 4740 were deposited in Japan Life Technology Institute Institute of Engineering and Industry Technology. It was deposited at the Institute of Food Industry Development on November 22, 1983, and the deposit numbers were CCRC 940 0 5 5 and 9 40 0 5 6. Example 5. Preparation of reconstituted human WS-4 antibody Repetitive rhythm of human WS-4 antibody Η chain V region The following design can encode DNA that reconstitutes the human WS-4 antibody Η chain V region. The known DNA sequences encoding FR1 ~ 3 of human antibody VDH26 and FR4 of human antibody 4 Β4, respectively, and then linked to the DNA encoding the CDRs of the mouse WS-4 antibody — chain V region, designed to encode the reconstruction of human The full-length DNA of the WS-4 antibody V chain. Secondly, the 5 &gt; -side and 3 / -side of the DNA sequence are added with the H i n dm recognition site / KOZAK common sequence and the B a mH I recognition site / cutting receptor sequence, so that they can be inserted into the Η E F expression vector. Divide the DNA sequence designed as above into approximately equal four oligonucleotides, and then use computer analysis to analyze the secondary structure in the oligonucleotides that may interfere with the collection of these oligonucleotides. Β Central of the Ministry of Economy The standardization bureau Shellfish Consumer Cooperative printed the four oligonucleotide sequences shown in sequences 3 2 to 35. These oligonucleotides have a length of 1 3 to 1 4 3 bases, and two adjacent oligonucleotides having 20 bases are regions that are mutually heavy. HF1 (sequence 32) and iHF3 (sequence 34) in the oligonucleotide have meaningful DNA sequences, and the other HF2 (sequence 33) and HF4 (sequence 35) have antisense DNA sequences. This is synthesized by an automatic DNA synthesis device (Applied Biosystems). The grades of oligonucleotide ° are applicable to the Chinese National Standard (CNS) A4 specifications (210X297 mm) -50-A7 B7 4 1 69SQ { Printed by the Standards Bureau Consumer Cooperatives V. Description of Invention (48) The method of assembling these four oligomeric light acids by the PC R method is shown in 囵 3. Each 100 ng of HF1 and HF2, HF3 and HF4 were combined and added to a PCR reaction solution containing 2,5 &quot; Pfu D N A polymerase with a final capacity of 9 8 // j ?. After the initial modification of 9 4 for 3 minutes, one cycle was applied at 94 ° C for 2 minutes, 55 ° C for 2 minutes, and 72 ° C for 2 minutes. After exchanging half the amount of the P C R reaction solution with each other, it was maintained at a constant temperature for the second cycle. After adding RVH5 &gt; primers (sequence 3 6) and RV Η 3 -primer (sequence 3 7) each as 10 Opmole as external primers, cover the PCR reaction solution with 5 0 &quot; $ mineral oil, on 9 4 After the initial modification at 3 ° C for 3 minutes, a cycle of 94 ° C for 1 minute, 55 ° C for 1 minute, and 72 ° C for 1 minute for 45 minutes, and finally incubation at 7 2 ° C for 10 minutes. The 45G base pair DNA fragment was constrained with 1.5% low melting point agarose gel, digested with Hindlll and BamHI, and then cloned into HEF-VH-g r 1, a HEF expression vector. After deciding the DNA sequence, it was named correctly containing the amino acid sequence that can encode the V region of the Η chain. The plastid of the DNA fragment was HEF — RVHa — g r 1. The amino acid sequence and base sequence of the fluorene chain V region contained in this plastid Η E F-R V — 1 — g r 1 are shown in SEQ ID NOs: 38. — The versions b, c, d, e, f, g, and h of the human WS-4 antibody WS chain V region were produced as follows. Version b (RVHb) uses LTW1 (Sequence 39) and LTW-2, which are designed to mutate the 47-position leucine to tryptophan (This paper size applies to China National Standards (CMS) A4 (210X 297) (%) Please read the note above and write this page A7 B7 Γ1 4169 € 0 V. Description of the invention (49) Sequence 4 〇) as the primer of the mutant, using plastid HEF — RVHa — gr1 as the template DNA, by Amplification by PCR method gave HEF_RVHb — gr 1. The amino acid sequence and base sequence of the fluorene chain V region contained in this plastid HEF — RVHb — g? * 1 are shown in sequence 41. The version c is based on the use of the original primer which can mutate glutamic acid at position 41 to proline mutation? 1 (sequence 42) and &lt; 3 but 卩 2 (sequence 4 3), plastid HEF ~ RVHa — g r 1 as template DNA, and amplified by PCR method to obtain plastid HEF — RVHc — gr 1. The amino acid sequence and base sequence of the fluorene chain V region contained in this plastid HEF — RVHc — g r 1 are shown in the sequence 4 4. Version d uses QTP 1 and QTP 2 as variant primers and plastid HEF — RVHb — g r 1 as template DNA to obtain plastid HEF — RVHd-gr 1. The amino acid sequence and fluorenyl sequence of the fluorene chain V region contained in this plastid HEF — RVHd — g r 1 are shown in the sequence 4 5. The printed version e of the Employees' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs is designed to mutate the alanine to proline mutant primers ATP1 (SEQ ID NO. 46) and ATP2 (SEQ ID NO. 47). -g? * l is used as template DNA amplification to obtain plastid HEF — RVHe_g r 1. The amino acid sequence and base sequence of the fluorene chain V region contained in this plastid HEF — RVHe_g r 1 are shown in sequences 48. Version f uses variant primers GTA1 (Sequence 4 9) and GTA2 (Sequence 50) designed to mutate glycine at position 4 to alanine. This paper is in accordance with Chinese National Standard (CNS) A4 (210X297 mm). 52 V. Description of the invention (50) A7 B7 Γ 418980, using the plastid HEF — RVHd-g τ · 1 as the template DNA, VHf — gr 1. In the sequence 5 1 represents the pyrene chain contained in VH f — gr 1. V-Amino Acid

Get the plastid HEF-R This plastid HEF-R sequence and base sequence. Version g is the variant primer LT 5 3), which is obtained from plastids Η DNA, and plastids Η indicates the amino acid sequence and base version contained in this plastids. H is obtained by using plastids HEF-R. Body EF — plastid HEF — RV column and base sequence. Designed to mutate the 6- and 7-position leucine into phenylalanine F 1 (sequence 5 2) and LTF 2 (sequence EF — RVHd — gr 1 as templates EF — RVHg — g − 1. 1. In sequence 5 4 EF-RVHg — The base sequence of the Η chain V region in gr 1. LTF 1 and LTF 2 are used as mutator primers, and VHb — gr 1 is used as the template DNA to amplify RVHh-gr 1. The sequence contained in H 5 — gr 1 is shown in sequence 5 5 For the amino acid sequence of the V region of the 阅读 chain, please read the note on the back again. On this page, printed by the Central Standards Bureau of the Ministry of Economic Affairs, Shellfish Consumer Cooperative, printed and reconstructed human WS _ 4 antibody The L chain V region is designed to encode human WS — 4 DNA of the V region of the antibody L chain. The DNA sequence encoding the FR of the human antibody RE I was designed to link to the DNA sequence of the CDR of the Iy bond V region of the mouse WS-4 antibody, which can encode the reconstructed human WS — 4 The full-length DNA of the L chain V region of the antibody. Secondly, one side and one side of this DNA sequence are added separately.

H ind ΙΠ recognition site / Κ 〇 Common sequence of ZAK and B am H This paper size is applicable to China National Standard (CNS) A4 (210X297 mm &gt; _ 53-at i 416980, B7 Printed by Staff Consumer Cooperative of Central Standards Bureau of Ministry of Economic Affairs Preparation of the invention (51) Recognition site / cutting receptor sequence 9 so that it can be inserted into the EF expression vector 0 The DNA sequence designed as above is divided into approximately equal four oligonucleotides, and the The analysis may have a secondary structure in the oligonucleotides that aggregate these oligonucleotides. The four oligonucleotide sequences are shown in the sequence 5 6 5 9 0 These oligonucleotides have 1 0 6 1 2 4 base lengths The adjacent two oligonucleosides have 1 9-23 bases which overlap each other. 0 LF 1 (sequence 5 6) 1 LF 3 {sequence 5 8) It has a phoneme D N A sequence, and other LF 2 (.Sequence 5 7) LF 4 (Sequence 5 9) has a negative D N A sequence. These oligonucleotides can be synthesized in the same way as ΗF 1 ~ 4 described above. The set (assemb I y) is a 9 8 U and P CR mixture containing 4 4 oligos each of 100 η g and 5 U of Amp! I T aq at 3 4 V for 3 minutes. The first modification is performed at 9 4 V 2 minutes, 5 5 ° C for 2 minutes, and 7 2 ° c for 2 minutes. Add RVL 5 primers (sequence 60) and RVL 3 / primer (sequence 6 1), each at 100 pm 〇1, and cover the PCR reaction solution with 50 μ 殳 mineral oil at 9 4 ° C as external primers. After the initial modification of 3 minutes, a cycle of 9 5 ° C for 1 minute, 7 5 5 ° C for 1 minute, and 7 2 V for 1 minute was applied for 4 5 cycles. Finally, the temperature was maintained at 7 2 V for 1 minute for 1 minute. 5% low melting point agarose gel was used to constrain 4 0 0 base pair D Ν Α fragments, digested with Η 1 η d III and B am Η I and then cloned into Η EF expression vector Η F F -VL — g κ 0 Determines the size of the DNA sequence paper to the Chinese National Standard (CNS) A4 specification (210X297 mm) _ 54-at! 4169 60 B7 Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of invention (52) Column 5 Name the plastid that correctly contains the DNA fragment that can encode the amino acid sequence of the L-chain V region as Η EF — RVL a — 8 κ 0. This plastid Ε EF-RVL a — SX contains the amine The amino acid sequence and base sequence are shown in the sequence 6 2 0 Version b (RVL b) uses FTY 1 (Sequence 6 3) and FTY 2 (Sequence 6) designed to mutate phenylalanine at position 71 to tyrosine 4) The plastid Η EF — RVL a-g κ as the template DNA was used as the mutant primer. The plastid was amplified by PCR. EF — RVL b-g X 0 represents this plastid in sequence 6 5 5 EF A RVL b — g K contains the amino acid sequence and test sequence of the L chain V Έ 0 is used to evaluate the antigen-binding activity of each chain of the reconstructed human WS-4 antibody 0 As described above, the human WS-4 antibody L is reconstructed Expression vector for chain version a Η EF — RVL a — SK and chimeric WS — 4 antibody 之 Expression vector for chain Η EF — C h WS 4 Η — gr 1 Simultaneously transfer infected C 0 S cells, recover and culture as described above After applying the supernatant, follow the method described in Example 4 ELISA above. The anti-tati body weight and antigen-binding activity were measured by the antibodies produced. The results are shown in Figure 40. Figure 4 shows the chimeric antibody (C h L / C h Η) as a positive control and the reconstituted L-bond and chimerism.抗体 It is confirmed that there is no difference in antigen binding between the antibodies formed by the chain (RVL a / C h Η) 0. It is also a performance carrier for evaluating the L bond of the chimeric W s 4 antibody Ε EF — C h WS 4 L Combination of 1 g K and reconstituted humanized WS-4 antibody Η chain a version &gt; transfer both to C 0 S cells at the same time &gt; apply to the obtained antibody test paper size according to the method described in Example 4 above ELISA Chinese National Standard (CNS) A4 specification (210X297 mm> _ η A7 B7 / 4 1 69 6) 5. Description of the invention (53) Determine the amount of antibody produced and antigen-binding activity. Results RVHa) did not see the original binding activity (such as the above-mentioned reconstructed human WS-4 anti-RVLa) line has resistance to chimeric WS-4, so each version of the other reconstruction chain is built to type WS- 4 Antibody L chain a version was transferred for evaluation. As a result, b, d'e, f, g, and the anti-system have the same degree of antigen-binding ability as that of the chimeric W / c h)) of the positive control, and a functional antigen-binding portion is formed in the human antibody.

This antibody (see Figure 4 somatic L chain a somatic L chain and reconstitution chain (RVL ch L /) 0 version (combined with each version and weight a)) Please refer to the note from the Central Bureau of Standards of the Ministry of Economic Affairs Consumption for work including the printed version except g (RVHg chL / chH) is shown to have antigen-binding activity (It can be seen that the functional antigen-binding site of an antibody with this (RVL a) and heavy (RVH g) can have no inferiority The amount of chimera generated. Secondly, the reconstructed RVL b) was used to simultaneously evaluate the reconstructed human type W. As a result, only the versions of the reconstructed Η chain with the reconstructed h S-4 antibody (ch L This shows that this combination can . But the amount of production is lower than the chimeric WS_4 antibody (lower. Moreover, the anti-system with the Η chain c version does not refer to Figure 5). Reconstructed human typed WS-4 antibody L chain a version built human typed The WS-4 antibody Η chain g version can be re-formed to have good antigen-binding properties. By simultaneously transferring infection to COS cells, human-typed WS-4 antibody (chL / chH) type WS-4 antibody L is transferred to Η Chain versions and CS-4 antibody L chain b version (human-typed WS — 4 antibody Η chain b version (〇 S cells, R V L b) chain g version Re-fill this page This page paper scale is applicable to China National Standard (CNS) A4 specifications (210X297 mmk% Chinese Ministry of Economic Affairs

X Consumer Cooperative 86; i .1 Γ 416960 Α7 h ΥΛ .χλ—-ϊ; .μ.; Β7 饨] especially j 5. Description of the invention (54) antibody (RVL b / RV Η S) shows no less than The degree of antigen binding of the chimeric WS-4 antibody (ch L / C h Η) of the positive control. In addition, in terms of production volume, except for the g version (RVH g), it is lower than the chimeric WS-4 antibody (ch L / C h Η) (see Figure 6). Two reconstituted humanized antibodies (RVL a / RVH g and RVL b / RVH g) that are not inferior to the chimeric WS-4 antibody (C h L / C h Η) in IL-8 binding activity, according to Example 4 ELISA The described method more accurately evaluates the binding activity * Results The chimeric WS-4 antibody (ch L / ch Η), RVL a / RV Η S antibody, and RVL b / RVH g antibody all showed the same degree of binding activity (see Figure 7) ). It can be seen that the anti-system with the reconstructed humanized WS-4 antibody L chain a version (RVL a) or b version (RVL b) and the reconstructed humanized WS-4 antibody Η chain g version (RV Η S) can be reformed. The functional antigen-binding site f, which has good antigen-binding properties, can be shown to be no less inferior to the chimeric WS-4 antibody (ch L / ch Η) by infecting C 0 S cells at the same time. The above coliforms with plastid HEF- RVL a — g λ are used as E. coli DH5 a (HEF-RVLa-g KK), and the coliforms containing plastid HEF -RVHg -g 71 are used as E. coli DH5a ( HEF— RVHg- gr 1) was deposited at the Institute of Biotechnology and Industrial Technology of the Japanese Institute of Industrial Technology under the International Depository Regulations of FERM BP-4438 and BP-444I under the Budapest Treaty on July 12, 1993 On November 22, 1983, it was deposited in the Q &quot; T'koukou Industry Development Research Institute 9 and the deposit numbers were CCRC 940 0 53 and 940 0 54. This paper size is applicable to China National Standard (CNS) A4 (210X297 mm) [77¾, 41 6960 V. Description of the invention (55 Reference Example 1 Making a fusion tumor WS-4 can produce anti-human IL-8 antibody fusion Tumors

• W Polyethylene glycol was used to fuse spleen cells of BALB / c mice immunized with human IL-8 and mouse bone marrow tumor cells P 3 X 6 3 — A g 8. 6 5 3 to make them. Screening was performed using the activity bound to IL-8 as an index to establish the fusion tumor WS-4. (K &lt; d, Y. C. et al., J. Iramunol. Methods, 149 * 227-235 »1 9 9 2) 0 v Industrial Applicability: The present invention can provide a reconstituted human antibody against human IL-8. The CD3R of the human anti- 鳢 V region in this antibody body was replaced by a mouse monoclonal antibody CDR against human IL-8. Most of the reconstructed human antibodies are derived from human antibodies, and CDR is originally low in antigenicity. Therefore, the reconstructed human anti-system of the present invention has low antigenicity to humans. Therefore, it can be expected to be used for medical therapy. Sequence List. 'Sequence Number: 1 Sequence Length: 4 0 Sequence Type: Nucleic Acid Strands: Single Strand Topology: Linear Sequence Type: Synthetic DNA-This paper applies Chinese National Standard (CNS &gt; Α4 Specification (210 × 297) (%) -58-(Please read the notes on the back before filling out this page) Order printed by the Intellectual Property Bureau Employee Consumer Cooperative of the Ministry of Economy A7 'X 416960 __ ^ _______ B7 _—_ V. Description of the invention (56) Sequence name: Μ Κ V 1 sequence: ACTAGTCGAC ATGAAGTTGC CTGTTAGGCT GTTCGTGCTG 40: sequence number: 2 sequence length: 3 9 sequence type: number of nucleic acid strands: single strand topology: linear sequence type: synthetic DNA sequence name: Μ Κ V 2 sequence: ACTAGTCGAC ATGGAGWCAG ACACACTCCT GYTATGGGT 39. S :: Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs Serial number: 3 Sequence length: 4 0 Sequence type: Nucleic acid strands: Single strand topology: Linear sequence type: Synthetic DNA Sequence name: Μ Κ V 3 sequence:.:. ¾ Called ACTAGTCGAC ATGAGTGTGC TCACTCAGGT CCTGGSGTTG 40 | This paper size applies to China National Standard (CNS) A4 specifications {210X (297 mm) -59-Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs a? Jf 416960 B7 V. Description of the invention (57) Serial number: 4 Sequence length: 4 3 Sequence type: Nucleic acid strand number: Single strand topology: Kind of linear sequence: Synthetic DNA Sequence name: Μ KV 4 Sequence: ACTAGTCGAC ATGAGGRCCC CTGCTCAGWT TYTTGGMWTC TTG 43 Sequence number: 5 Sequence length: 4 0 Sequence type: Nucleic acid strand number: Single strand topology: Linear sequence type: Synthetic DNA sequence name: Μ Κ V 5 sequence: ACTAGTCGAC ATGGATTTWC AGGTGCAGAT TWTCAGCTTC 40 sequence number: 6 sequence length: 3 7 sequence type: number of nucleic acid strands: single stock paper size, using China National Standard (CNS) A4 size (210X297 mm) ) &Lt; Please read the notes on the back before writing this page)

A7 ('4169 ^ 0 _____ Β7__ 5. Description of the invention (58) Topology: Linear sequence type: Synthetic DNA Sequence name: Μ KV 6 Sequence: ACTAGTCGAC ATGAGGTKCY YTGYTSAGYT YCTGRGG ^ 37 Sequence number: 7 Sequence length: 4 1 Sequence type : Number of nucleic acid strands: Single-strand topology: Linear sequence type: Synthetic DNA Sequence name: ΜΚ V 7 Sequence: ACTAGTCGAC ATGGGCWTCA AGATGGAGTC ACAKWYYCWG G 41 丨 Central Bureau of Standards, Ministry of Economic Affairs, Zhengong Consumer Cooperative Co., Ltd. Printed sequence number: 8 Sequence Length: 4 1 Sequence type: Nucleic acid strand number: Single-strand topology: Linear sequence type: Synthetic DNA Sequence name: Μ Κ V 8 Sequence IJ: 61 This paper size applies to China National Standard (CNS) Α4 specification (210 × 297 (Mm) I 416960 A7, __ B7 V. Description of the invention (59) ACTAGTCGAC ATGTGGGGAY CTKTTTYCMM TTTTTCAATT G 4: Sequence number: 9 Sequence length: 3 5 Sequence type: Nucleic acid strand number: Single strand topology: Linear sequence type: Synthetic DNA sequence name: Μ KV 9 sequence:

IcTAGTCGAC ATGGTRTCCW CASCTCAGTT CCTTG 35 Printed by the Consumer Standards Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs Serial number: 1 0 Sequence length: 3 7 Sequence type: Number of nucleic acid strands: Single strand topology: Linear sequence type: Synthetic DNA Sequence name: MKV 1 0 Sequence: ACTAGTCGAC ATGTATATAT GTTTGTTGTC TATTTCT 37 Sequence number: 1 1 Sequence length: 3 8 62 This paper size applies to Chinese National Standard (CNS) A4 (210X 297 mm) f 416960 A7 'B7 Central Bureau of Standards of the Ministry of Economic Affairs-Industry Printed by Consumer Cooperatives

V. Description of the invention (60) Sequence type: fucidic acid strands: single strand topology: linear sequence type: synthetic DNA Sequence name: M KV 1 1 Sequence: ACTAGTCGAC ATGGAAGCCC CAGCTCAGCT TCTCTTCC 38; Sequence number: 1 2 Sequence Length: 2 7 Sequence type: Nucleic acid strand number: Single strand topology: Linear sequence type: Synthetic DNA Sequence name: Μ KC Sequence: GGATCCCGGG TGGATGGTGG GAAGATG 27 Sequence number: 1 3 Sequence length: 3 7 Sequence type: Nucleic acid strand Number: Single-strand topology: Linear sequence Type: Synthetic DNA This paper uses Chinese National Standard (CNS) A4 specifications (210X297 mm) _ 416960 A7 B7 Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs

5. Description of the invention (61) Sequence name: M Η V 1 Sequence: ACTAGTCGAC ATGAAATGCA GCTGGGTCAT STTCTTC Sequence number: 1 4 Sequence length: 3 6 Sequence type: Nucleic acid strand number: Single strand topology: Linear sequence type: Synthetic DNA Sequence name: Μ Η V 2 Sequence: ACTAGTCGAC ATGGGATGGA GCTRTATCAT SYTCTT Sequence number: 1 5 Sequence length: 3 7 Sequence type: Nucleic acid strand: Single strand topology: Linear sequence type: Synthetic DNA Sequence name: Μ Η V 3 Serial: ACTAGTCGAC ATGAAGWTGT GGTTAAACTG GGTTTTT 37 36 37 Please note

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The paper size of the edition applies to the Chinese national standard (CNS &gt; Α4 specification (210 father 297 mm)-64-i 41S9S0 Α7 Β7 V. Description of the invention (62) Sequence number: 1 6 Sequence length: 3 5. Sequence type: Number of nucleic acid strands: single strand topology: linear sequence type: synthetic DNA sequence name: Μ Η V 4 sequence: ACTAGTCGAC ATGRACTTTG GGYTCAGCTT GRTTT 35

Employees' Cooperative Cooperative I of the Central Bureau of Standards of the Ministry of Economic Affairs Serial number: 1 7 Sequence length: 4 0 Sequence type: Nucleic acid strand number: Single strand topology: Linear sequence type: Synthetic DNA Sequence name: M Η V 5 Sequence: ACTAGTCGAC ATGGACTCCA GGCTCAATTT AGTTTTCCTT 40 sequence number: 1 8 sequence length: 3 7 sequence type: number of nucleic acid strands: single stock paper wave scale applicable to China National Standard (CNS) A4 specifications (210X2,97 Gong) _still_ A7 B7 f 416980 Economy Printed by the Central Bureau of Standards, Shellfish Consumer Cooperative, V. Description of the Invention (63) Topology: Linear sequence type: Synthetic DNA Sequence name: M Η V 6 Sequence: ACTAGTCGAC ATGGCTGTCY TRGSGCTRCT CTTCTGC Sequence number: 1 9 Sequence length: 3 6 Sequence type: Number of nucleic acid strands: Single-strand topology: Linear sequence type: Synthetic DNA Sequence name: M Η V 7 Sequence: ACTAGTCGAC ATGGRATGGA GCKGGRTCTT TMTCTT Sequence number: 2 0 Sequence length: 3 3 Number: single strand topology: linear sequence type: synthetic DNA sequence name: Μ Η V 8 sequence: (Please read the note on the back first Matters need to be written on this page) -5 This paper scale is applicable to the Chinese National Standard (CNS) A4 specification (2 [0X2.97 mm). The Ministry of Economic Affairs ordered the Central Bureau of Standards and Staff to print the A7 f 4169 60 B7 V. Description of the invention (64) ^ CTAGTCGAC ATGAGAGTGC TGATTCTTTT GTG 33 sequence number: 2 1 sequence length: 4 0 sequence type: number of nucleic acid strands: single strand topology: linear sequence type: synthetic DNA sequence name: Μ Η V 9 Sequence: ACTAGTCGAC ATGGMTTGGG TGTGGAMCTT GCTATTCCTG 40 Sequence number: 2 2 Sequence length: 3 7 Sequence type: Nucleic acid strand number: Single strand topology: Straight-chain! Sequence type: Synthetic DNA Sequence name: M Η V 1 0 Sequence: ACTAGTCGAC ATGGGCAGAC TTACATTCTC ATTCCTG 37 serial number: 2 3 serial length: 3 8 This paper size is applicable to the Chinese national standard (〇 呢) person 4 specifications (210 乂 2.97 mm) _6 _ (Please read the precautions on the back and write on this page first )

, 1T A7 B7 f * 416SS0

V. Description of the invention (65) Sequence type: Nucleic acid strand number: Single strand topology: Linear sequence type: Synthetic DNA Sequence name: M Η V 1 1 Sequence: ACTAGTCGAC ATGGATTTTG GGCTGATTTT TTTTATTG Sequence number: 2 4 Sequence length: 3 7 Sequence type: Nucleic acid strand number: Single-strand topology: Straight-chain sequence 'Column type: Synthetic DNA Sequence name: Μ Η V 1 2 Sequence: ACTAGTCGAC ATGATGGTGT TAAGTCTTCT GTACCTG 38 37 Please read the precautions for backing Sr again / Ordered by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs

Sequence number: 2 5 Sequence length: 2 8 Sequence type: Nucleic acid strand number: Single-strand topology: Linear sequence type: Synthetic. DNA This paper size is applicable to Chinese National Standard (CNS) A4 specification (210X297 mm) Member of the Central Bureau of the Ministry of Economic Affairs, X. Consumption cooperative printed A7 '1' 41 B7 'V. Description of the invention (66)

Sequence name: M H C Sequence: 28! GGATCCCGGG CCAGTGGATA GACAGATG 1

Sequence number: 2 6 Sequence length: 3 8 2 Sequence type: Nucleic acid strand number: Double-strand topology: Linear sequence type: c DNA Sequence name: WS 4 VL Origin ^ Biological name: Pure germline name of mouse direct origin : PXJC-WS4—VL Features: 1..60 signal peptide 61 .. 382 mature peptide sequence! ATG AGT GTG CTC ACT CAG GTC CTG GGG TTG CTG CTG CTG TGG CH 45

Met Ser Val Leu Thr Gin Val Leu Gly Leu Leu Leu Leu Trp Leu -20 -15 -10 ACA GGT GCC AGA TGT GAC ATC CAG ATG ACT CAG TCT CCA GCC TCC 90 |

Thr Gly Ala Arg Cys Asp lie Gin Met Thr Gin Ser Pro Ala Ser -5 -11 5 10 j! CTA TCT GCA TCT GTG GGA GAA ACT GTC ACC ATC ACA TGT CGA GCA 135; Leu Ser Ala Ser Val Gly Glu Thr Val Thr lie Thr Cys Arg Ala; 15 20 25 · j AGT GAG ATT ΑΠ TAG ACT TAT TTA GCA TGG TAT CAG CAG AAA CAG 180

Ser Glu lie lie Tyr Ser Tyr Leu AU Trp Tyr Gin Gin Lys Gin 30 35 40 (Precautions for reading on the back of this page are written on this page) Ding,-'b. This paper size applies Chinese National Standard (CNS) A4 specifications ( 210 parent 297 mm) _ 69 A7 | 4169S® B7 Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs V. Invention Description (67) GGA AAA TCT CCT CAG CTC CTG GAT TAT AAT GCA AAA ACC TTA GCA Gly Lys Ser Pro Gin Leu Leu Val Tyr Asn Ala Lys Thr Leu Ala 45 50 55 GAT GGT GTG TCA TCA AGG TTC AGT GGC AGT GGA TCA GGC ACA CAG Asp Gly Val Ser Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Gin 60 65 70 τπ TCT CTG CGG ATC AGC AGC CTG CAG CCT GAA GAT TTT GGG AGT Phe Ser Leu Arg He Ser Ser Leu Gin Pro Glu Asp Phe Gly Ser 75 80 85 TAT TAC TGT CAA CAT CAT ΤΠ GGT TTT CCT CGG ACG TTC GGT GGA Tyr Tyr Cys Gin His His Phe Gly Phe Pro Arg Thr Phe Gly Gly 90 95 100 GGC ACC AAG CTG GAA CTC AAA C Gly Thr Lys Leu Glu Leu Lys 105 Sequence number: 2 7 Sequence length: 4 2 4 Sequence type: Nucleic acid strand number: Double-strand topology : Linear sequence type: c DNA sequence Name: WS4VH Origin biological name: Mouse direct origin Pure germline name: PUC_WS 4 — VH Features: 1..57 signal peptide 58..424 mature peptide sequence 25 70 60 38 (Please read the notes on the back first ^: write (This page) This paper size applies to China National Standard (CNS) A4 (210X297 mm) · 70-V. Description of invention (68) A7 B7 4 1 69 6Ό, 'ATG MG TTG TGG TTA MC TCG GTT TTT CTT GTG ACA CTT ΤΤΑ AAT Met Lys Leu Trp Leu Asa Trp Val Phe Leu Val Thr Leu Leu Asn -19 -15 -10 -5 GGT ATC CAG TGT GAG GTG AAA CTG GTG GAG TCT GGA GGA GGC HG Gly lie Gin Cys Glu Val Lys Leu Val Glu Ser Gly Giy Gly Leu -11 5 10 ATA CAG CCT GGG GAT TCT CTG AGA CTC TCC TCC GGT ACC TCT GGG lie Gin Pro Gly Asp Ser Leu Arg Leu Ser Cys Val Thr Ser Gly 15 20 25 TTC ACC TTC AGT GAT TAC TAC CTG AGC TGG GTC CGC CAG CCT CCA Phe Thr Phe Ser Asp Tyr Tyr Leu Ser Trp Val Arg Gin Pro Pro 30 35 40 GGA AAG GCA 0Ή GAG TGG GTG GGT CTC ΑΠ AGA AAC AAA GCC AAT Gly Lys Ala Leu Glu Trp Val Gly Leu lie Arg Asn Lys Ala Asn 45 50 55 GGT TAC ACA AGA G AG TAC AGT GCA TCT GTG AAG GGT CGG TTC ACC Gly Tyr Thr Arg Glu Tyr Ser Ala Ser Val Lys Gly Arg Phe Thr 60 65 70 ATC TCC AGA GAT GAT TCC CAA AGC ATC CTC TAT CTT CAA ATG AAC lie Ser Arg Asp Asp Ser Gin Ser lie Leu Tyr Leu Gin Met Asn 75 80 85 ACC CTG AGA GGT GAG GAC AGT GCC ACT TAT TAC TGT GCA CGA GAG Thr Leu Arg Gly Glu Asp Ser Ala Thr Tyr Tyr Cys Ala Arg Glu 90 95 100 Central Ministry of Economy AAC TAT AGG TAC GAC GTA GAG CTT GCT TAC TGG GGC CAA GGG ACT Asn Tyr Arg Tyr Asp Val Glu Leu Ala Tyr Trp Gly Gin Gly Thr 105 05 115 CTG GTC ACT GTC TCT GCA G Leu Val Thr Val Ser Ala 120 45 90 135 180 225 II ί 270! 315 360: i 405. 424: (Please read the note on the back first and write this page)

I equipment -------- IT li ^ · This paper size is applicable to Chinese National Standard (CNS) A4 specification (210X297 mm) -71 κ Β7 I1 4 169 6v0, V. Description of invention (69) Serial number: 2 8 Sequence length: 3 4 Sequence type: Nucleic acid strand number: Single strand topology: Linear sequence type: Synthetic DNA Sequence name: ch VL rear primer sequence ACAAAGCTTC CACCATGAGT GTGCTCACTC AGGT 34 Sequence number: 2 9 Sequence length: 3 7 Sequence type : Nucleic acid strand number: Single strand topology: Linear sequence type: Synthetic DNA Sequence name: ch V ΗBack primer sequence GATAAGCTTC CACCATGAAG TTGTGGTTAA ACTGGGT 37 Sequence number: 3 0 Sequence length: 3 7 Sequence type: Nucleic acid strand number: Single The paper size of the share capital is applicable to the national standard of the country (&lt; ^ 45) 8-4 specification (210 mm 297 mm) -72- (Please read the precautions on the back #, fill out this page), -a Employees of the Central Standards Bureau of the Ministry of Economic Affairs Printed by Consumer Cooperatives Printed by Consumers' Cooperatives, Central Bureau of Standards, Ministry of Economic Affairs

i '4 169CT Α7 Β7 V. Description of the invention (7〇) Topology: Linear sequence type: Synthetic DNA Sequence name: chVL front primer sequence CTTGGATCCA CTCACGTTTG AGTTCCAGCT TGGTGCC 37: Sequence number: 3 1 Sequence length: 3 7 Sequence type : Nucleic acid strands: Single strand topology: Linear

Sequence type: synthetic DNA Sequence name: chVH forward primer sequence GTCGGATCCA CTCACCTGCA GAGACAGTGA CCAGAGT 37: sequence number: 3 2 sequence length: 1 3 7 sequence type: number of nucleic acid strands: single strand topology: linear sequence type: synthetic DNA sequence Name: HF 1 sequence (please read the precautions on the back before filling this page) --------------- The size of the paper is applicable to China National Standard (CNS) Α4 specification (2 丨 〇X 297 mm) _ _ a? 4 1 6960 _B7 _.__ V. Description of the invention (Ή) 50 I 1〇〇! 137;

^ AAGCTTCCA CCATGGAGTT TGGGCTGAGC TGGGTTTTCC TTGTTCCTAT TTTAAAGGGT GTCCAGTGTG AAGTGCAGCT GTTGGAGTCT GGGGGAGGCT TGGTCCAGCC TGGGGGTTCT CTGAGACTCT CATGTGC Sequence number: 3 3 Sequence length: 1 type: strand type: Nucleic acid sequence type: Nucleic acid: 2 sequence 50 100 143

j; CACTGTACT CTCTTGTGTA ACCATTGGCT TTGTTTCTAA TGAGACCCAC CAACTCTAGC CCTTTCCCTT GAGCTTGGCG GACCCAGCTC AGGTAGTAAT CACTGAAGGT GAATCCAGAG GCAGCACATG AGAGTCTCAG AGA sequence number: 3 4 sequence length: 1 1 3 sequence type: straight strand sequence DNA: single strand type: single number: ： HF 3 series This paper size applies to China National Standard (CNS &gt; Α4 size (210X297mm) A7 B7

41696Q Printed by the Shellfish Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs 5. Description of the invention (72) TACACAAGAG AGTACAGTGC ATCTGTGAAG GGCAGACTTA CCATCTCAAG 50 II AGAAGATTCA AAGAACACGC TGTATCTGCA AATGAGCAGC CTGAAAACCG 100 | AAGACTTGGC CGT 113 | i Sequence number: 3 5 Sequence length: 1 5 Type: Nucleic acid strand number: Single strand topology: Linear sequence type: Synthetic DNA Sequence name: HF 4-sequence 'TCGGATCCAC TCACCTGAGG AGACGGTGAC CAGGGTTCCC TGGCCCCAGT 50 AAGCAAGCTC TACGTCGTAG CGATAGTTCT CTCTAGCACA GTAATACACG 100 GCCAAGTCTT CGGTTTT 6 sequence length: 117 3 7 Sequence type: Number of nucleic acid strands: Single-strand topology: Types of linear sequences · Synthetic DNA Sequence name: RVH5 &lt; Primer sequence The size of this paper applies to China National Operation (CNS) A4 specification (2! 〇 × 297 mm ) -75, please read the precautions before

Order 416960 A7 B7 Printed by the Central Laboratories Consumers Cooperative of the Ministry of Economic Affairs. 5. Description of the invention (73) liATAAGCTTC CACCATGGAG TTTGGGCTGA GCTGGGT 37 Sequence number: 3 7 Sequence length: 4 4 Sequence type: Nucleic acid strands: Single strand topology: Type of linear sequence: Synthetic DNA Sequence name: RV Η 3 ^ Primer sequence JGTCGGATCCA CTCACCTGAG GAGACGGTGA C ^ 31 Sequence number: 3 8 Sequence length: 424 Sequence type: Number of nucleic acid strands: Double-strand topology: Type of linear sequence: Synthetic DNA sequence name: RV Η a Origin biological name: Mouse and human direct origin Pure germline name: Η EF-RVH a-gr 1 Amino acid — 1 9-— 1: Please read the note S above «Write this page 1 The size of this paper is in accordance with the Chinese National Standard (CNS) A4 (210X29 &quot;? mm) _ 7 Fifth, the description of the invention (74) A7 B7 4 1 6960 Amino acid 1-3 0 FR 1 Amine Acid 3 1 —3 5 CDR 1 amino acid 3 6 -4 9 FR 2 amino acid 5 0 —6 8 CDR 2 amino 6 9 -1 0 0: F R 3 amino acids 1 0 1 - 1 1 1 • C D R amino acid 1 1 2 - 1 2 2: F R 4 sequences is read CAUTIONS read back of the

Printed by ATG GAG TTT GGG CTG AGC TGG GTT TTC CTT GTT GCT ATT TTA AAG Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala lie Leu Lys -19 -15 -10 -5 GGT GTC CAG TGT GAA GTG CAG CTG TTG GAG TCT GGG GGA GGC TTG Gly Val Gin Cys Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu _1 1 5 10 GTC CAG CCT GGG GGT TCT CTG AGA CTC TCA TGT GCT GCC TCT GGA Val Gin Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 15 20 25 TTC ACC TTC AGT GAT TAC TAC CTG AGC TGG GTC CGC CAA GCT CAA 'Phe Thr Phe Ser Asp Tyr Tyr Leu Ser Trp Val Arg Gin Ala Gin 30 35 40 GGG AAA GGG CTA GAG TTG GTG GGT CTC ATT AGA AAC AAA GCC AAT Gly Lys Gly Leu Glu Leu Val Gly Leu lie Arg Asn Lys Ala Asn 45 50 55 GGT TAC ACA AGA GAG TAC AGT GCA TCT GTG AAG GGC AGA CTT ACC Gly Tyr Thr Arg Glu Tyr Ser Ala Ser Val Lys Gly Arg Leu Thr 60 65 70 ATC TCA AGA GAA GAT TCA AAG AAC ACG CTG TAT CTG CAA ATG AGC lie Ser Arg Glu Asp Ser Lys Asn Thr Leu Tyr Leu Gin Met Ser 75 80 85 AGC CTG AM ACC GAA GAC TTG GCC GTG TA T TAC TGT GCT AGA GAG Ser Leu Lys Thr Glu Asp Leu Ala Val Tyr Tyr Cys Ala Arg Glu 90 95 100 360 Order A7 .ί &quot; 4ΐβΒδϊ _____ Β7_ V. Description of Invention (75) AAC TAT CGC TAC GAC GTA GAG CTT GCT TAC TGG GGC CAG GGA ACC 405

Asn Tyr Arg Tyr Asp Val GIu Leu Ala Tyr Trp Gly Gin Gly Thr 105 110 115 CTG GTC ACC GTC TCC TCA G 424 Leu Val Thr Val Ser Ser 120 Sequence number: 3 9 Sequence length: 3 4 Sequence type: Number of nucleic acid strands: Single strand topology: Linear sequence type: Synthetic DNA Sequence name: LTW 1 Sequence GGCTAGAGTG GGTGGGTCTC ATTAGAAACA AAGC 34 Sequence number: 4 0 Sequence length: 3 6 Printed by the Central Standards Bureau of the Ministry of Economic Affairs, Masonry Consumer Cooperatives Sequence type: Nucleic acid stock Number: single strand topology: straight chain

Sequence type: Synthetic D N A Sequence name: LTW2 sequence

I CAGACCCACC CACTCTAGCC CTTTCCCTTG AGCTTG 3 &amp; This paper size is applicable to Chinese National Standard (CNS) Α4 size (210X297 mm) _ 78. Α7 Β7 Γ 416960 Printed by Bayer Consumer Cooperative, Central Bureau of Standards, Ministry of Economic Affairs 5. Description of invention (π) Sequence number: 4 1 Sequence length: 4 2 4 Sequence type: Nucleic acid strand number: Double-strand topology: Linear sequence type: Synthetic DKfA Sequence name: RVH b Origin biological name: mouse and human g Direct origin pure germline Name: Η EF Amino acid-1 9 Amino acid 1 Amino acid 31 Amino acid 36 Amino acid 50 Amino acid 6 9 Amino acid 10 Amino acid 11 Sequence RVH b-gr 1 —1: leader 3 5 4 9 6 8 FR 1 CDR 1 FR 2 CDR 2 ~ 10 0: FR 3 1- 111: CDR 3 2- 122: FR 4 ATG GAG TTT GGG CTG AGC TGG GTT TTC CTT GTT GCT Απ ΠΑ AAC 45 Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala He Leu Lys ~ 19 -15 -l〇-5 GGT GTC CAG TGT GAA GTG CAG CTG HG GAG TCT GGG GGA GGC TTG 90 Gly Vai Gin Cys Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu ..... This paper size applies Chinese National Standard (CNS) A4 (210X297mm) _ 79-f &quot; 4 1 6960 V. Description of invention (77) GTC CAC CCT GGG GGT TCT CTG AGA CTC TCA TGT GCT GCT TCT GGA 135

Val Gin Pro 0iy Gly Ser ieu Arg Leu Ser Cys Ala Ala Ser Gly 15 20 25 HC ACC TTC AGT GAT TAC TAC CTG AGC TGG GTC CGC CAA GCT CAA 180

Phe Thr Phe Ser Asp Tyr ITyr ieu. Ser. Trp Vai rg Gin Aia ό 30 35 40 GGG AAA GGG CTA GAG TGG GTG GGT CTC ATT AGA AAC AAA GCC AAT 225

Gly Lys Gly Leu Glu Trp Val Gly Leu lie Arg Asn Lys Ala Asn 45 50 55 GGT TAC ACA AGA GAG TAC AGT CCA TCT GTG AAG GGC AGA CH ACC 270

Gly Tyr Thr Arg Glu Tyr Ser Ala Ser Val Lys Gly Arg Leu Thr 60 65 70 ATC TCA AGA GAA GAT TCA AAG AAC ACG CTG TAT CTG CAA ATG AGC 315 lie Ser Arg Glu Asp Ser Lys Asn Thr Leu Tyr Leu Gin Met Ser 75 80 85 AGC CTG AAA ACC GAA GAC TTG GCC GTG TAT TAC TGT GCT AGA GAG 360

Ser Leu Lys Thr Glu Asp Leu Ala Val Tyr Tyr Cys Ala Arg Glu 90 95 100 i AAC TAT CGC TAC GAC GTA GAG CTT GCT TAC TGG GGC CAG GGA ACC 405)

Asn Tyr Arg Tyr Asp Val Glu Leu Ala Tyr Trp Gly Gin Gly Thr 105 110 115 CTG GTC ACC GTC TCC TCA G 424

Leu Val Thr Val Ser Ser 120 Serial number printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs • 4 2 Sequence length; 3 2 Sequence type • Number of fusic acid single-strand topology Topology Science Synthetic DNA Sequence name QTP 1 Sequence The scale of this paper is in accordance with China National Standards (CNS) A4 (210X 297 mm) -go-416960 A7 B7 Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (78) TGGGTCCGCC AAGCTCCAGG GAAAGGGCTA GA} 32 sequence Number: 4 3 Sequence length: 3 2 Sequence type: Nucleic acid strand number: Single strand topology: Linear sequence type: Synthetic DNA Sequence name: Q T 2 Sequence TCTAGCCCTT TCCCTGGAGC TTGGCGGACC CA 32 Sequence number: 4 4 Sequence length: 4 2 4 Sequence type: Nucleic acid strand number: Double-strand topology: Linear sequence type • Synthetic D ΝΑ Sequence name: RV Η c Origin biological name: Mouse and human direct origin pure germline name: Η EF-RVH c -gr 1 Amino acid-1 9 ——1: 1 eader This paper size applies Chinese National Standard (CNS) Α4 size (210 X 297 mm) _ 8 workers Please read the precautions before writing this page A7 B7 ^ 416960 V. Description of the invention (79) amino acid amino acid amino acid amino acid amino acid amino acid amino acid sequence 1-3 1-3 3 6 4 9 5 0-68 6 9-10 10 1 — 1 112 — 1 FR 1 CDR 1 FR 2 CDR 2: FR 3 1: CDR 3 2: FR 4 Read first note l l; AG TTT Met Glu Phe GGT CTC CAG Gly Val Gin GGG CTG AGC TGG GTT Gly Leu Ser Trp Vai -15 TGT GAA GTG CAG CTG Cys Glu Val Gin Leu TTC CTT GTT GCT ATT πΑ MG Phe Leu Val Ala lie Leu Lys -10 -5 45 Items 1 Write this page

-1 GTC CAG CCT Val Gin Pro TTC ACC TTC Phe Thr Phe GGG AAA GGG Gly Lys Gly GGT TAC ACA Gly Tyr Thr Printed by ATC TCA AGA lie Ser Arg AGC CTG AM Ser Leu Lys GGG GGT TCT CTG AGA Gly Gly Ser Leu Arg 15 AGT GAT TAC TAC CTG Ser Asp Tyr Tyr Leu 30 CTA GAG HG CTG GGT Leu Glu Leu Val Gly 45 AGA GAG TAC AGT GCA Arg Glu Tyr Ser Ala 60 GAA GAT TCA AAG AAC Glu Asp Ser Lys Asn 75 ACC GAA GAC TTG GCC Thr Glu Asp Leu Ala 90 TTG GAG TCT GGG GGA GGC TTG Leu Glu Ser Gly Gly Gly Leu 5 10 CTC TCA TGT GCT GCC TCT GGA Leu Ser Cys Ala Ala Ser Gly 20 25 AGC TGG GTC CGC CAA GCT CCA Ser Trp Val Arg Gin Ala Pro 35 40 CTC ATT AGA AAC AAA GCC AAT Leulie Arg Asn Lys Ala Asn 50 55 TCT GTG AAG GGC AGA CH ACC Ser Val Lys Gly Arg Leu Thr 65 70 ACG CTG TAT CTG CAA ATG AGC Thr Leu Tyr Leu Gin Met Ser 80 85 GTG TAT TAC TGT GCT AGA GAG Val Tyr Tyr Cys Ala Arg Glu 95 100 90 135 180 225 270 315 3G0 (2! 0 &gt; &lt; 297 mm)- 82-41 «9β〇 B7 Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs

A7 丨 ‘‘ 4 1 6960 B7 V. Description of the invention (81) ATG GAG Τπ GGG CTG AGC TGG GTT TTC CTT GTT GCT ATT ΠΑ AAG Ί5:

Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala lie Leu Lys) -19 -15 -10 -5 i GGT GTC CAG TGT GAA GTG CAG CTG TTG GAG TCT GGG GGA GGC TTG 90

Gly Val Gin Cys Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu -11 ... 5 10 GTC CAG CCT GGG GGT TCT CTG AGA CTC TCA TGT GCT GCC TCT GGA 135!

Val Gin Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 15 20 25 TTC ACC TTC ACT GAT TAC TAC CTG AGC TGG GTC CGC CAA GCT CCA 180

Phe Thr Phe Ser Asp Tyr Tyr Leu Ser Trp Val Arg Gin Ala Pro 30 35 40 GGG AAA GGG CTA GAG TGG GTG GGT CTC ATT AGA AAC AAA GCC AAT 225

Gly Lys Gly Leu Glu Trp Val Gly Leu lie Arg Asn Lys Ala Asn 45 50 55 GGT TAC ACA AGA GAG TAC AGT GCA TCT GTG AAG GGC AGA CTT ACC 270

Gly Tyr Thr Arg Glu Tyr Ser Ala Ser Val Lys Gly Arg Leu Thr 60 65 70 ATC TCA AGA GAA GAT TCA AAG AAC ACG CTG TAT CTG CAA ATG AGC 315 lie Ser Arg Glu Asp Ser Lys Asn Thr Leu Tyr Leu Gin Met Ser 75 80 85 AGC CTG AAA ACC GAA GAC TTG GCC GTG TAT TAC TGT GCT AGA GAG 360

Ser Leu Lys Thr Glu Asp Leu Ala Val Tyr Tyr Cys Ala Arg Glu 90 95 100 AAC TAT CGC TAC GAC GTA GAG CTT GCT TAC TGG GGC CAG GGA ACC 405:

Asn Tyr Arg Tyr Asp Val Glu Leu Ala Tyr Trp Gly Gin Gly Thr 105 no ns CTG GTC ACC GTC TCC TCA G 424

Leu Val Thr Val Ser Ser 120 84-This paper size applies to Chinese National Standards (CNS) A4 (210 X 2_97 mm) _ A7 B7 f &lt;, 4 1 6960 V. Description of the invention (82) Serial number: 4 6 Serial Length: 2 6 Sequence type: Number of nucleic acid strands: Single strand topology: Linear sequence type: Synthetic DNA Sequence name: ATP 1 Sequence TGGGTCCGCC AACCTCCAGG GAAAGG: Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs

Sequence number: 4 7 Sequence length: 2 6 Sequence type: Nucleic acid Number of strands: Single strand Topology: Straight chain Sequence type: Synthetic D N A Sequence name: A T P 2 Sequence IJ

CCTTTCCCTG GAGGTTGGCG GACCCA Sequence number: 4 8 Sequence length: 4 2 4 Sequence type: Nucleic acid strands: Double equity Paper size Applicable to China National Standard (CNS) A4 specifications (210X'297 mm) 85-A7 B7 ί · 41 6960 5 Explanation of the invention (83) Topology: Linear sequence type: Synthetic DNA Sequence name: RVH e Origin biological name: Mouse and human direct origin Pure germline name: HE F — RV Η e — gr 1 Amino acid- 1 9-— 1 1 eacer Amino acid 1-3 0 FR 1 Amino acid 3 1-3 5 CDR 1 Amino acid 3 6-4 9 FR 2 Amino acid 5 0-6 8 CDR 2 Amino acid 6 9-1 0 0 ； FR 3 Amino acid 1 0 1 — 1 1 1 • C DR Amino acid 1 1 2 _ 1 2 2 • FR 4 (Please read the precautions on the back before you ^;: Write this page ) Order the printed sequence of the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economy

ATG GAG TTT GGG CTG AGC TGG GTT TTC CTT GTT GCT ATT TTA AAG

Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala lie Leu Lys -19 -15 _l〇

GGT GTC CAG TGT GAA GTG CAG CTG TTG GAG TCT GGG GGA GGC TTG

Gly Val Gin Cys Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu -11 5 10

GTC CAG CCT GGG GGT TCT CTG AGA CTC TCA TGT GCT GCC TCT GGA

Val Gin Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly | 15 20 25

TTC ACC TTC AGT GAT TAC TAC CTG AGC TGG GTC CGC CAA CCT CCA

Phe Thr Phe Ser Asp Tyr Tyr Leu Ser Trp Val Arg Gin Pro Pro 30 35 40 This paper size is free from the Chinese National Standard (CNS) A4 specification (210X297 mm) * 00 5. Description of the invention (84) A7 'f &quot; 4 I 6960 B7 GGG AAA GGG CTA GAG TGG GTG GGT CTC Απ AGA AAC AAA GCC AAT Gly Lys Gly Leu Glu Trp Val Gly Leu lie Arg Asn Lys Ala Asn 45 50 55 GGT TAC ACA AGA GAG TAC ACT GCA TCT GTG AAG GGC AGA CTT ACC Gly Tyr Thr Arg Glu Tyr Ser Ala Ser Val Lys Gly Arg Leu Thr 60 65 70 lie W 5AT fAG iAC ACG CTG TAT CTG CAA ATG AGC He ber Arg Glu Asp Ser Lys Asn Thr Leu Tyr Leu Gin Met Ser 75 80 85 · Ifr LeS tS fAC irTG STf ΪΑΤ TAC TGT GCT AGA GAG ber Leu Lys Thr Glu Asp Leu Ala Val Tyr Tyr Cys Ala Arg Glu 90 95 100 AAC TAT CGC TAC GAC GTA GAG CH GCT TAC TGG GGC CAG GGA ACC Asn Tyr Arg Tyr Asp Val Glu Leu Ala Tyr Trp Gly Gin Gly Thr 105 HO 115 CTG GTC ACC GTC TCC TCA G Leu Val Thr Val Ser Ser 120 Central Standards Bureau of the Ministry of Economic Affairs-Industrial and Consumer Cooperation Du printed serial number: 4 9 sequence length: 2 9 sequence Type: Nucleic acid strands: Single Topology: Linear Sequence Type: Synthetic DNA Sequence Name: G T A 1 Sequence &lt; iAAGCTCCAG GGAAAGCGCT AGAGTGGGT Sequence Number: 5 0 (Please read the notes on the back before filling this page)

This paper size applies to China National Standard (CMS &gt; A4 size (210X297 mm) _ π fv 4 1 69 60 Β7

Employees' Cooperative Cooperative Seal of the Central Bureau of Standards, Ministry of Economic Affairs I. Description of the invention (85) Sequence length: 2 9 Sequence type: Number of nucleic acid strands: Single strand topology: Linear sequence type: Synthetic DNA Sequence name: GTA 2 Sequence ACCCACTCTA GCGCTTTCCC TGGAGCTTG sequence number: 5 1 sequence length: 4 2 4 sequence type: number of nucleic acid strands: double-strand topology: linear sequence type: synthetic DNA sequence name: RVH f origin biological name: mouse and human direct origin purebred Series name: Η EF-RVH f-gr 1 Amino acid-1 9—— 1: Leader Amino acid 1-3 0: FR 1 Amino acid 3 1-3 5: CDR 1 Amino acid 3 6-4 9: FR 2 This paper size is in accordance with Chinese National Standard (CNS) A4 specification (21 〇 × 29 * 7 mm) _ ___ B7, f :, 4 1 6960 V. Description of the invention (86) Amino acid 5 0-68 • CDR 2 amino acid 6 9 — 10 0: FR 3 amino acid 10 1-1 1 1 CDR 3 amino acid 112-1 2 2: FR 4 sequence

ATG GAG ΤΠ GGG CTG AGC TGG GH m CTT GTT GCT ATT TTA AAG

Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala lie Leu Lys; -19 -15 -10 -5 j GGT GTC CAG TGT GAA GTG CAG CTG TTG GAG TCT GGG GGA GGC TTG |

Gly Val Gin Cys Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Gly Leu: -11 5 i〇

GTC CAG CCT GGG GGT TCT CTG AGA CTC TCA TGT GCT GCC TCT GGA

Val Gin Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly, 15 20 25

TTC ACC TTC AGT GAT TAC TAC CTG AGC TGG GTC CGC CAA GCT CCA

Phe Thr Phe Ser Asp Tyr Tyr Leu Ser Trp Val Arg Gin Ala Pro 30 35 40

GGG AAA GCG CTA GAG TGG GTG GGT CTC ATT AGA AAC AAA GCC AAT

Gly Lys Ala Leu Glu Trp Val Gly Leu lie Arg Asn Lys Ala Asn 45-50 55

GGT TAC ACA AGA GAG TAC AGT GCA TCT GTG AAG GGC AGA CTT ACC

Gly Tyr Thr Arg Glu Tyr Ser Ala Ser Val Lys Gly Arg Leu Thr 60 65 70

ATC TCA AGA GAA GAT TCA AAG AAC ACG CTG TAT CTG CAA ATG AGC lie Ser Arg Glu Asp Ser Lys Asn Thr Leu Tyr Leu Gin Met Ser 75 80 85

AGC CTG AAA ACC GAA GAC TYQ GCC GTG TAT TAC TGT GCT AGA GAG

Ser Leu Lys Thr Glu Asp Leu Ala Val Tyr Tyr Cys Ala Arg Glu 90 95 100

AAC TAT CGC TAC GAC GTA GAG CTT GCT TAC TGG GGC CAG GGA ACC

Asn Tyr Arg Tyr Asp Val Glu Leu Ala Tyr Trp Gly Gin Gly Thr 105 110 115 CTG GTC ACC GTC TCC TCA G Leu Val Thr Val Ser Ser 120 This paper size applies to the Chinese National Standard (CNS) A4 (210 X 297 mm) ) _ Gg Printed at I '* 41 6960 B7 by the Consumer Co-operation of the Central Standards Bureau of the Ministry of Economic Affairs V. Description of the Invention (87) · Serial number: 5 2 Sequence length: 2 3 Sequence type: Nucleic acid strands: Single strand topology : Linear Sequence Type: Synthetic DNA Sequence Name: LTF 1 Sequence

GTGAAGGGCA GATTTACCAT CTC sequence number: 5 3 sequence length: 2 3 sequence type: nucleic acid strand number: single strand topology: straight chain sequence type: synthetic DNA sequence name: L T F 2 sequence

GAGATGGTAA ATCTGCCCTT CAC Sequence number: 5 4 Sequence length: 4 2 4 Sequence type: Nucleic acid strands: Double-equity Paper size Applicable to China National Standard (CNS) Α4 specification (2 丨 0X297 mm) --------- '3pack. 丨 | C (please read the notes on the back before filling out this page) Order five. Description of the invention (δ8) Topology: Linear sequence type: Synthetic DNA Sequence name: RVHg Origin biological name: Mouse And human direct origin pure germline name amino acid amino acid amino acid amino acid amino acid amino acid amino acid amino acid sequence I * 4 1 6960 HEF — RVHf —gr 1 —1 9 — — 1 :! eader 1 — 30 3 1 — 35 3 6 — 49 5 0-6 8

F R 1 C D R F R 2 C D R

6 9-1 0 0: FR 3 1 0 1-1 1 1: CDR 1 1 2 1 2 2: FR 4 (谙 Please read the notes on the back before writing this page) Printed by a cooperative

ATG GAG ΤΓΤ GGG CTG AGC TGG GTT HC CTT GH GCT ΑΠ ΠΑ MG ket Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala lie Leu Lys -19 -15 -10 -5

GGT GTC CAG TGT GAA GTG CAG CTG TTG GAG TCT GGG GGA GGC TTG

Gly Val Gin Cys Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu -11 5 10

GTC CAG CCT GGG GGT TCT CTG AGA CTC TCA TGT GCT GCC TCT GGA

Val Gin Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 15 20 25 This paper is quickly used in China National Standards (CNS) A4 specifications (21 OX 2.97 mm) _ gi Printed by the Staff Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs A7 r 4 t 6960 B7 V. Description of the invention (89)

i TTC ACC TTC ACT GAT TAC TAC CTG AGC TGG GTC CGC CAA GCT CCA 丨 Phe Thr Phe Ser Asp Tyr Tyr Leu Ser Trp Val Arg Gin Ala Pro 30 35 40

GGG AAA GGG CTA GAG TGG GTG GGT CTC ATT AGA AAC AAA GCC AAT

Gly Lys Gly Leu Glu Trp Val Gly Leu lie Arg Asn Lys AJa Asn 45 50 55 GGT TAC ACA AGA GAG TAC ACT GCA TCT GTG AAG GGC AGA TTT ACC Gly Tyr Thr Arg Glu Tyr Ser Ala Ser Val Lys Gly Arg Phe Thr 60 65 70

ATC TCA AGA GAA GAT TCA AAG AAC ACG CTG TAT CTG CAA ATG AGC lie Ser Arg Glu Asp Ser Lys Asn Thr Leu Tyr Leu Gin Met Ser 75 80 85: AGC CTG AAA ACC GAA GAC TTG GCC GTG TAT TAC TGT GCT AGA GAG Ser Leu Lys Thr Glu Asp Leu Ala Val Tyr Tyr Cys Ala Arg Glu 90 95 100

'AAC TAT CGC TAC GAC GTA GAG CTT GCT TAC TGG GGC CAG GGA ACC; Asn Tyr Arg Tyr Asp Val Glu Leu Ala Tyr Trp Gly Gin Gly Thr; 105 110 115 I CTG GTC ACC GTC TCC TCA G Leu Val Thr Val Ser Ser 120 Sequence number: 5 5 Sequence length: 4 2 4 Sequence type: Nucleic acid strand number: Double-strand topology: Linear · Sequence type: Synthetic DNA Sequence name: RVH h Origin biological name: Mouse and human direct origin purebred Department name: HEF — RVHh — gr 1 Amino acid-1 9——1: leader This paper size applies to China National Standard (CNS &gt; A4 specification (2! 0X297 mm) _ 92 ------- -------- ΐτ, m,--I (Please read the notes on the back before 4iv · script 写) A7 B7 416960 V. Description of the invention (90) Printed by the Consumers' Cooperative of the Central Bureau of Quasi-Ministry of Economic Affairs Amino acid 1 -3 0-FR 1 Amino acid 3 1 -3 5: CDR 1 Amino acid 3 6 -4 9 FR 2 Amino acid 5 0-6 8; CDR 2 Amino acid 6 9 -1 0 0: FR 3 amino acid 1 0 1-1 1 1-CDR amino acid 1 1 2 — 1 2 2 FR 4 sequence 'ATG GAG ΤΓΤ GGG CTG AGC TGG GTT HC CTT GTT GCT ATT TTA AAG Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala He Leu Lys -19 -15 -10 -5 GGT GTC CAG TGT GAA GTG CAG CTG TTG GAG TCT GGG CGA GGC HG Gly Val Gin Cys Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Gly leu -11 5 l〇GTC CAG CCT GGG GGT TCT CTG AGA CTC TCA TGT GCT GCC TCT GGA Val Gin Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Giy 15 20 25 TTC ACC TTC ACT GAT TAC TAC CTG AGC TGG GTC CGC CAA GCT CAA Phe Thr Phe Ser Asp Tyr Tyr Leu Ser Trp Val Arg Gin Ala Gin 30 35 40 GGG AAA GGG CTA GAG TGG GTG GGT CTC ATT AGA AAC AAA GCC AAT Gly Lys Gly Leu Glu Trp Val Gly Leu lie Arg Asn Lys Ala Asn 45 50 55 GGT TAC ACA AGA GAG TAC AGT GCA TCT GTG AAG GGC AGA ΤΓΤ ACC Gly Tyr Thr Arg Glu Tyr Ser Ala Ser Val Lys Gly Arg Phe Thr 60 65 70 1 ATC TCA AGA GAA GAT TCA AAG AAC ACG CTG TAT CTG CAA ATG AGC 'lie Ser Arg Glu Asp Ser Lys Asn Thr Leu Tyr Leu Gin Met Ser; 75 80 85 AGC CTG AAA ACC GAA GAC GAC HG GCC GTG TAT TAC TGT GCT AGA CAG Ser Leu Lys Thr Glu Asp Leu Ala Val Tyr Tyr Cys Ala Arg Glu I 90 95 100

This paper size applies to Chinese National Standard (CNS) A4 specification (2〗 0X297 mm) -9J J Pack — (Please read the precautions on the back before filling this page), \ = 口-.) ¾ A7 r 41 6960 B7 V. Description of Invention (91) AAC TAT CGC TAC GAC GTA GAG CH GCT TAC TGG GGC CAG GGA ACC Asn Tyr Arg Tyr Asp Val Glu Leu Ala Tyr Trp Gly Gin Gly Thr HO 115 (Please read the notes on the back first (Write this page) CTG GTC ACC GTC TCC TCA G Leu Val Thr Val Ser Ser 120 Sequence number: 5 6 Sequence length: 1 2 4 Sequence type: Nucleic acid strand number: Single strand topology: Linear sequence type: Synthetic DNA sequence Name: LF 1 Sequence

TTGAAGCTTC CACCATGGGA TGGAGCTGTA TCATCCTCTT CTTGGTAGCA ACAGCTACAG GTGTCCACTC CGACATCCAG ATGACCCAGA GCCCAAGCAG CCTGAGCGCC AGCGTAGGTG ACAG Printed AN 2 D 2 7 2 into the F 5 1 acid chain Straight L ..... ： Single: Class title Long type ： Student name list Column list Column list Column order Column order Array order Scroll order Scroll paper size Chinese National Standard (CNS) A4 specification (210X297 mm) 94 41 6960 Α7 Β7 Ministry of Economic Affairs Printed by the Central Sample Bureau, Shellfish Consumer Cooperative Co., Ltd. 5. Description of the invention (Θ2) sequence [gcattgtaga tcagcagctt tggagccttt cctggcttct gctggtacca TGCTAAATAA CTGTAAATAA TCTCGCTTGC TCGACAGGTG ATGGTCACTC TGTCACCTAC GCTGGCGCTC AG 1 Sequence number: 5 Sequence number: 1 Single-strand topology: Linear sequence type: Synthetic DNA Sequence name: LF 3 sequence ^ AGCTGCTGAT CTACAATGCA AAAACCTTAG CAGATGGAGT GCCAAGCAGA TTCAGCGGTA GCGGTAGCGG TACCGACTTC ACCTTCACCA TCAGCAGCCT CCAGCCAGAG GACATCGCTA C .... Sequence number: 5 9 sequence type: Number of acid strands: Single strand topology: Linear sequence type: Synthetic DNA Sequence name: LF 4 Precautions for prior notice Binding for each paper size Applicable to China National Standard (CNS) M size (210x297 mm) 95-Economy A7 '4 1 69 60 B7 printed by the Consumer Cooperatives of the Ministry of Standards of the People's Republic of China V. Invention Description (93) Sequence

GTAGGATCCA CTCACGTTTG ATTTCGACCT TGGTCCCTTG GCCGAACGTC!

IcGAGGAAAAC CAAAATGATG TTGGCAGTAG TAGGTAGCGA TGTCCTCTGG CTGGAG sequence number: 6 0. Sequence length: 2 0 sequence type: nucleic acid strand number: single strand topology: straight chain sequence type: synthetic DNA sequence name: R V L 5 — sequence

TTGAAGCTTC CACCATGGGA sequence number: 6 1 sequence length: 2 0 sequence type: nucleic acid strand number: single strand topology: straight chain sequence type: synthetic DNA sequence name: R V L 3 &gt; sequence

GTAGGATCCA CTCACGTTTG This paper size applies to Chinese National Standard (CNS) + A4 specification (210X297 mm) _ {Please read the precautions on the back before filling this page)

Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs A7 Γ 41 6960 __'_ B7_ V. Description of the invention (94) Serial number: 6 2 Sequence length: 3 7 9 Sequence type: Nucleic acid strands: Double strand topology: Straight-chain Sequence type: Synthetic DNA Sequence name: RVL a Origin biological name: Mouse and human direct origin Pure germline name: HEF-RVLa-g γ Amino acid i 9——1: leader Amino acid 1 — 23: FR1 Amino acid 24 — 34: CDR1 Amino acid 35-49: FR2 Amino acid 50 — 56: CDR2 Amino acid 57-88: FR3 Amino acid 89-97: CDR3 Amino acid 98-107: FR4 Sequence ATG GGA TGG AGC TGT ATC ATC CTC TTC TTG GTA GCA ACA GCT ACA Met Gly Trp Ser Cys lie lie Leu Phe Leu Val Ala Thr Ala Thr -19 -15 -10 -5 I -------- install ----- ---- Order ------ ^. Line (read the precautions on the back and then write this page on the wall) This paper size is applicable to China National Standard (CNS) A4 (210X297 mm) 97 A7 f 41 69 60 B7 V. Description of the invention (9S) GGT GTC CAC TCC GAC ATC CAG ATG ACC CAG AGC CCA AGC AGC CTG Gly Val His Ser Asp lie Gin Met Thr Gin Ser Pro Ser Ser Leu! -1 1 5 10: AGC GCC AGC GTA GGT GAC AGA GTG ACC ATC ACC TGT CGA GCA AGC 1 Ser Ala Ser Val Gly Asp Arg Val Thr lie Thr Cys Arg Ala Ser 15 20 25 GAC ATT ATT TAC AGT TAT TTA GCA TGG TAC CAG CAG AAG CCA GGA Glu He lie Tyr Ser Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly 30 35 40 AAG GCT CCA AAG CTG CTG ATC TAC AAT GCA AAA ACC ΠΑ GCA GAT Lys Ala Pro Lys Leu Leu lie Tyr Asn Ala Lys Thr Leu Ala Asp 45 50 55 GGA GTG CCA AGC AGA TTC AGC GGT AGC GGT AGC GGT ACC GAC TTC Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 60 65 70 i ACC TTC ACC ATC AGC AGC CTG CAG CCA GAG GAC ATC GCT ACC TAC! Thr Phe Thr lie Ser Ser Leu Gin Pro Glu Asp lie Ala Thr Tyr 75 80 85 TAC TGC CAA CAT CAT TTT GGT TTT CCT CGG ACG TTC GGC CAA GGG Tyr Cys Gin His His Phe Gly Phe Pro Arg Thr Phe Gly Gin Gly 90 95 100 ACC AAG GTC GAA ATC AAA C Thr Lys Val Glu lie Lys 105 (Please read the notes on the back first to write this page).

'IT Printing by Central Standards Bureau, Ministry of Economic Affairs, Consumer Cooperatives

Sequence number: 6 3 Sequence length: 3 8 Sequence type: Nucleic acid strand number: Single-strand topology: Linear sequence type: Synthetic DNA Sequence name: FTY 1 Sequence AGCGGTAGCG GTACCGACTA CACCTTCACC ATCAGCAG S This paper is in accordance with Chinese national standards ( CNS) A4 specification (210X297 mm) η〇 &quot; 98 ί * B960 Μ B7 Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (96) Sequence number: 6 4 Sequence length: 3 8 Sequence type: Nucleic acid Number of strands: single strand topology: linear sequence type: synthetic DNA sequence name: FTY 2 sequence iTGCTGATGG TGAAGGTGTA GTCGGTACCG CTACCGCT sequence number: 6 5 sequence length: 3 7 9 sequence type: nucleic acid strand number: double strand topology: straight Type of chain sequence: Synthetic DNA Sequence name: RVL b Originating biological name: Mouse and human direct origin Pure germline name: HEF — RVLb — gx Amino acid — 1 9 — — Γ: leader Amino acid 1 — 2 3 : FR 1 Amino acid 24 — 34: CDR1 (Please read the precautions on the back before writing this page) Binding. The size of the paper is applicable to the Chinese National Standard (CMS) Α4 Grid (210X297 mm) V. Description of the invention (97) A7 B7 416960 Amino acid 3 5--49: FR 2 Amino acid 5 0--5 6: CDR 2 Amino acid 5 7 ~ -8 8: FR 3 Amine Jun 8 9--9 7: CDR 3 Amino acid 9 8--1 0 7: FR 4 For the sequence, please refer to the printing of ATG GGA TGG AGC TGT ATC ATC CTC TTC TTG GTA GCA ACA GCT ACA Met Gly Trp Ser Cys lie lie Leu Phe Leu Val Ala Thr Ala Thr -19 -15 -10 -5 GGT GTC CAC TCC GAC ATC CAG ATG ACC CAG AGC CCA AGC ACC CTG Gly Val His Ser Asp lie Gin Met Thr Gin Ser Pro Ser Ser leu -11 5 10 AGC GCC AGC GTA GGT GAC AGA GTG ACC ATC ACC TGT CGA GCA AGC Ser Ala Ser Val Gly Asp Ar.g Val Thr lie Thr Cys Arg Ala Ser 15 20 25 GAG ATT ATT TAC ACT TAT TTA GCA TGG TAC CAG CAG AAG CCA GGA Glu He lie Tyr Ser Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly 30 35 40 AAG GCT CCA AAG CTG CTG ATC TAC AAT GCA AAA ACC TTA GCA GAT Lys Ala Pro Lys Leu Leu He Tyr Asn Ala Lys Thr Leu Ala Asp 45 50 55 GGA GTG CCA AGC AGA TTC AGC GGT AGC GGT AGC GGT ACC GAC TAC Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr 60 65 70 ACC HC ACC ATC ACC AGC CTC CAG CCA GAG GAC ATC GCT ACC TAC Thr Phe Thr lie Ser Ser Leu Gin Pro Glu Asp lie Ala Thr Tyr 75 80 85 TAC TGC CAA CAT CAT TTT GGT TTT CCT CGG ACG TTC GGC CAA GGG Tyr Cys Gin His His Phe Gly Phe Pro Arg Thr Phe Gly Gin Gly i 90 95 100; ACC AAG GTC GAA ATC AAA C Thr Lys Val Glu lie Lys! 105 τ6 Precautions fL · Paperback Dimensions Applicable to China National Standards (CNS) A4 Specification (210 X 297 mm) _ 100

Claims (1)

A8 B8 C8 D8 416960 VI. Application for Amendment of Patent Scope S / 89. 8- 4Wi Patent Application No. 83110990 Chinese Application for Amendment of Patent Scope (Please read the notes on the back before filling this page) August 89, Republic of China Amendment 1. The light chain (L chain) variable region (V region) of a mouse monoclonal antibody against human interleukin-8 (IL-8) has the following amino acid sequence shown in sequence 26: Asp Us Gin Met Thr Gin Ser Pro Ala. Ser Leu Ser Ala Ser Vai 1 5. 10 15 Gly Glu Thr.; Yal Thr lie Thr Cys Arg Ala Ser'Giu lie lie Tyr 20 25 .30 Ser Tyr Leu Ala Trp Tyr Gin Gin Lys Gin Gly--Lys Ser Pro Gin t 35 40 45 Leu Leu Va.1 Tyr Asn Ala Lys Thr ^ Leu Ala Asp Gly Yal Ser Ser 50 '; 55 60 Arg Phe Ser Gly Ser Gly Ser Gly Thr Gin Phe Ser Leu Arg lie 65 70 75 Ser Stir Leu Gin Pro Glu Asp Phe Gly Ser Tyr Tyr Cys Gin His Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs 80 35 90 Ris Phe Gly Phe Pro Arg Thr Phe Cly Gly Gly Thr Lys Leu GU '95 100 105 Leu Lys 2. A mouse monoclonal antibody against human IL-8 Chain (H chain V region, which has the following amino acid sequence shown in sequence 27:. This paper size applies to the China National Standard (CNS) A4 specification (21〇x 297 mm) § V | * 4 \ 69 60 VI. Application scope Clu Vil Lys Leu Val Glu Ser Gly Gly Gly leu lie Gin Pro Cly 1 5 10 15 Asp Ser Leu Arg Leu Ser Cys Yal Thr Ser Gly Phe Thr Phe Ser 20 II 25 30 Asp Tyr Tyr Leu Ser Trp Val Arg Gin Pro Pro Gly Lys Ala Leu 35 '40 45 Glu Trp Val Gly Leu ile Arg Asn Lys Ala Asn Gly Tyr Thr Arg 50 55 60 Gl.u Tyr Ser AJa Ser Val Lys Gly Arg Phe Tfir Ile Ser Arg Asp 65 70 75 Asp Ser Gin Ser Ile Leu Tyr Leu Gin Met Asn Thr Leu Arg Gly 80. 85 90 Glu Asp Ser Ala Thr Tyr Tyr Cys Ala Ar? Giu Asn Tyr Ar Xuan Ding 95 100 i ° 5 Asp Val Glu Leu Ala Tyr Trp Gly Gin Gly Thr Leu Val Thr Val 110 115 120 Ser Ala 3. A chimeric antibody L chain characterized by containing a human antibody 1-chain fixed region (C region) and a mouse monoclonal antibody against human IL-8 In the L chain of the resulting chimeric antibody, the aforementioned mouse [chain 7 region is the following amine shown in the order% .26 Acid Sequence: The elimination of this scale with Zhang Yin 3 Ξ mark this house (CNS) A4 size (210 * 297 male S) (please, "read the Notes on the back of Benedict reloading the page) Install -------- Order --------- ^ u · el t 4 1 6960 C.8 D8 VI. Patent Application Range Asp lie Gln. Met Thr Gin Ser Pro Ala Ser Leu Ser AU Ser Vat L 5 10. 15 QlY Giu Thr Val Thr lie Thr Cys Arg Ala Ser Glu He lie Tyr 20 25 30 Ser Tyr Leu Ala. Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro Gin 35 40 '45 Leu Leu Val Tyr Asn Ala lys Thr leu Ala Asp Gly Val Ser Ser 50 55 60 Arg Plae Ser Gly Ser Cly Ser Gly Thr Gin Pbe Ser Leu hrg lie. 65 70 75 Ser Ssr Leu Gin Pro Glu Asp Phe Gly Ser Tyr Tyr Cys Gin His 80 85 9〇His Phe Gly Phe Pro Ar? Thr Phe Gly Gly Gly Thr Lys Leu Glu 95, 100 l〇5 Leu Lys Printed by the Employee Consumption Cooperative of the Intellectual Property Office of the Ministry of Economic Affairs ^ (Please read the precautions on the back before filling in this page 4.) A chimeric antibody H chain, which is characterized in that the chimeric antibody Η chain formed by the 单 bond V region of the mouse monoclonal antibody against human i L-8 containing the 11 bond C region of the human antibody is as described above. The V region of the mouse bond is the following amino acid sequence which is different from the sequence 2 7: 'Ben. ¾ ¾ feet. Supporting SS Tsukamaki (C &gt; «TS) A ·! Gauge * grid (210 X 2 97 issued) A8B8C8D8 4 ο 6 9 6 Consumption cooperation on off-work and consumption of the Intellectual Property Bureau of the Ministry of Economic Affairs 矣 Application scope Glu Val Lys Leu Val Glu Ser Gly Gly Gly Leu lie Gin Pro Gly .1 5 '10 15? .Sp Ser Leu Arg Leu Ser Cys Val Thr Sir Gly Phe Thr Phe Ser 20: '25 30 Asp Tyr Tyr Leu Ser Trp Yal Arg CIn Pro Pro Gly Lys Ala Leu 3S 40 45 Glu Trp Val Gly teu lie Arg Asn Lys Ala Asn Gly Tyr Thr Arg so 55 .........- 60 Glu Tyr Ser Ala Ser Yal Lys Gly Ars Phe Thr lie Ser Arg Asp 65 70 75 Asp Ser Gin Ser lie Leu Tyr Leu Gin Met Asn Thr Leu Ar.g Gly 80 .. 85 90 Glu Asp Ser Ala Thr Tyr Tyr Cys Ala Arg .Glu Asn Tyr kZ Tyr 95 100 i ° 5 Asp Vil Glu Leu Ala Tyr Trp Gly Gin Gly Thr leu Va Thr Vai 110 115 120 Ser AU. ·-5.—Species A chimeric antibody is characterized by containing: (1) an L chain V region of a mouse monoclonal antibody containing a human antibody L chain fixing region (C region) and an anti-human IL-8; and (2) containing Human antibody Η chain C region and anti-human I + mouse monoclonal antibody Η chain V region, the resulting Η chain; in the resulting chimeric antibody, the aforementioned mouse L chain V region contains a sequence The following amino acid sequences of column 2 6 # $: (Please read the notes on the back before filling this page) ^ * * 111111 — &lt; cr · Paper: ¾ standard applicable S3 standard (Ci \ TS) A4 (210x297 male) ― 4-A8 B8 C8 D8 416960 6. Application scope of patent isp He Gin Met Thr Gin Ser Pro Mi Ser Leu Ser Ala Ser Val 1 5 10 15 Cly GU Thr Vai Thr He Thr Cys Arg Ala Ser Glu He lie Tyr 20. 25 30 Ser Tyr Leu Ala Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro Gin 3S 40 45 Leu Leu Val Tyr Asn AU Lys Thr leu Ala 'Asp Cly Vai Ser Ser 50 55,. 60 Ar? Phe Ser Gly Ser Gly Ser Gly Thr Gin Pile Ser Leu Arg lie 65 70 75 Ser Ser Leu Gin Pro Glu Asp Phe Gly Ser Tyr Tyr Cys Gin His 80 85 90 Eiis Phe Gly Phe Pro Arg Thr Phe Gly Cly Gly Thr Lys Leu Glu 95 100 105 Leu Lys. \ And the aforementioned mouse H chain V region has the following amino acid sequence described in sequence ^ 7 Glu Yal Lv-s Ls Val Glu Ser Gly Gly Gly Leu lie Gin Pro Gly 1 5 l〇I5 Asp Ser Leo. Λ: Leu Ser Cys Yal Thr Ser Gly Phe Thr Phe Ser '20 25 30 •. Ser Trp Val Arg Gin Pro Pro Gly Lys Ala Leu 35 40 45 Leu 丨 Ie Arg Asn Lys All.Asn Gly Tyr Thr Arg 50 55 -60 Use the paper to the full scale and use the order 舀 Suka Kita Jun (CNS) A4 specifications (210 x 297 public release), please pay attention to the items on JH n's back Pages Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs, India, Asp Tyr Tyr L Glu Trp Vil A8 B8 C8 D8 110, eu 4 1 69 60:., Patent application scope GU Tyr Ser Ai Ser Val L / s Gly Ar «Phe Thr Ile Ser Arg Asp 65 70 75 Asp Ser CltL S: Ile Lea Tyr Leu Gin Met Asn Thr Leu krz Cly 80. 85 90 Glu Asp Ser Ai Ttir T.yr ΠΓ CH AU Arg Glu Asn T.yr Arg Tyr 95 100 105 Asp Val G; i in Ala Tyr TrP Gly Gin Gly'Thr Leu Val Thr Val .115, 'i20 Ser Ala 6, a complementarity determining region of the L bond V region of a mouse monoclonal antibody against human IL-8 ) 'It is the amino acid sequence CDR 1 shown below; Arg Ala Ser Glu Ile Tyr Ser Tyr Ala' CDR 2; Asn Ala Lys Thr Leu Ala Asp CDR 3; Gin His His Phe Gly Phe Pro Arg Thr — CDR of the H-bond V region of a mouse monoclonal antibody against human IL.-8, which has the amino acid sequence shown below: CDR 1; Asp Tyr Tyr Leu Ser CDR 2; Leu'Ile Arg Asn Lys Ala Asn Gly Tyr Thr Arg Glu Tyr Ser Ala Ser Val Lys GlY CDR 3; Glu Asn Tyr Arg Tyr Asp Val Glu L eu Ala • Tyr 8.—Reconstitution of anti-human I. L-8 antibodies to human antibodies L ^ V Paper: ¾ ft. Applicable t gg due to grazing sheep (CNS) A4 specification (21〇χ 297 gong), y 、-、、. ----------- α 装 -------- Order --------- (please note ¾ on the back of Μίί before filling in this f) Consumers ’cooperation with the Intellectual Property Bureau of the Ministry of Economic Affairs, India 5Λ A8 B8 C8 D8 416960 VI. Patent application area, which is characterized by containing (1) the frame structure area CFR of the human antibody L chain V area), and (2) for anti-human The CDR of the L chain V region of the mouse monoclonal antibody of IL-8 is formed by the veshaped human antibody L chain V region of the human IL-8 antibody, wherein the aforementioned CDR has the following amino acid sequence: CDR 1; Ars Ala Ser G [u IU He Tyi · 'Ser Tyr Leu Ala CDR 2; Asn Ala lys Thr Leu Ala Asp, CDR 3; Gin His His Phe Gly Plie Pro Arg Thr and the aforementioned FR is from the human antibody RE I. 9. The reconstructed human antibody L chain V region according to item 8 of the patent application scope, wherein the L chain V region is an amino acid sequence having RVL a as shown below. (Please read the notes on the back before filling this page) Printed on paper by the Consumers' Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs. The paper size is applicable to the Chinese National Standard (CNS) A ·! Specifications (210 X 297). 7 VI. Application for patent scope 88 r 4 1 6960 C8 * D8 1 2 12345678901234567890123 3 45678901234 4 567890123456789 RVLa DIQMTQSPSSLSASVGDRVTITC RASEIIYSYLA WYQQKPGKAPKLLIY FR1 CDR1 .V FR2 5 6 7 8 9 (Please read the notes on the back before filling this page) Consumer Co-operation of the Intellectual Property Bureau of the Ministry of Economic Affairs, printed by Du 0123456 78901234567890123456789012345678 901234567 • -r, RVLa NAKTLAD GVPSRFSGSGSGTDFTFTISSLQPEDIATYYC QHHFGFPRT CDR2 FR3 CM3 10. 8901234567- RVLa FGQGTKVEIK FR4 · Chinese Standards (Applicable to China) 210 x 297 public love) -8-A8 BS C8 D8 f- 41 69 60 VI. Application for patent scope 1 0 · —Reconstructed human antibody H 鐽 ▽ region of an anti-human 1 L-8 antibody, which is characterized by containing (1 ) FR of the human antibody Η chain V region, and &lt; 2) to human IL-8 mouse antibody Η chain V region CDRs formed 'for human IL-8 antibody reconstruction of human antibody Η chain V region , Where the aforementioned CDRs have the following amino acid sequences: CDR 1; Asp Tyr Tyr Leu Ser CDR 2; Leu lie Arg Asn Lys Ala Asn Gly Tyr Thr Arg Glu • &gt; Tyr Ser Ala Ser Yal lys Gly CDR 3: Glu Asn Tyr Are Tyr Asp Val Glu Leu Ala Tyr and FR1, 2, 3 of the aforementioned FR are derived from the human antibody VDH2 6, and FR4 is derived from the human antibody 4B4. 1 1. The reconstituted human antibody Η chain V region according to item 10 of the patent application scope, wherein the Η chain V region has the following RVHa, RVH b, RVHc. RVH d 'RVH e, RVHf' RVHg or RVHh Amino acid sequence · Please read Note I on the back first, ★ This page IIIII Orders: Intellectual Property Bureau of the Ministry of Economic Affairs § Industrial Consumers Cooperative Cooperative Seal ¥-Rule Paper. Ϊ́ Standard S Inge 8 00 8 8 A ^ CD 1- 41 6960 67890123456789 VI. Application for Patent Range 1 2 3 123456789012345678901234567890 12345 RVHa ~ h EVQLLESGGCLVQPGGSLRLSCAASGFTFS DYYLS FR1., CDR1 5 6 012ABC3456739012345 (Please read the notes on the back before filling out this page) Printed by the Consumer Affairs Cooperative of the Ministry of Intellectual Property Bureau of the Ministry of Justice-RVHa RVHb RVHc RVHd RVHe RVHf RVHg RVHQG LVYG -VIRG-LIRNK —- · — — pp —, ---- FR2 CDR2 This paper has been used in the standard a S home standard (G \ S) A4 specification (210 X 297 gong) -10-6. Scope of patent application RVHa RVHb RVHc RVHd RVHe RVHf RVHg RVHh RVHa ～ h A8 B8 C8 D8! '4 1 6960 7 8 9 67890123456789012ABC345678901234 100 567890ABC12 RLTISREDSKNTLYLQMSSLKTEDLAVYYCAR ENYRYDVELAY &lt; Please read the first page of MR3, please read the pre-reading of MR3, please read the back of this page for more information.- 3-Read the first page of CDR, please read the first page and read the first page and read the third page. Please read the first page and read the first page of the CDR. 3-5. Please read the first page of the CDR and read it first. Please read the first page of MR3, please read the first page of the CDR. Please read the first page on the back page of the RVHc RVHd. FR4 This paper is a long-range application of the Chinese SS family fS standard (CNS) A4 specification (21 0 χ 297 males) 11-A8 B8, _S f 4 1 6960 6. Application for patent scope 12-a reconstituted human antibody l chain against human I L_8, which is characterized by containing (1) the human antibody L chain C region And (2) the L chain V region formed by the L chain CDRs of the human monoclonal antibody L chain FR and the anti-human IL-8 mouse monoclonal antibody, wherein the human antibody l chain C region is a human C / C region, human antibody L chain FR is from re I, the L chain CDR has the amino acid sequence shown below: CDR 1; Arg Ala Ser'Glu He Tyr'Ser Tyr Leu Ala CDR 2; Asn Ala Lys Thr Leu Ala Asp CDR 3; Gin His His Phe Gly Phe Pro Arg Thr (Please read the note on the back first? Please fill in this page again.) Chain L Sequencer Acid Anti-Amine Amines Human 1 Construction a Heavy L V Item R 2 1 Shown under the rule of Fan Ruli, please specify the string V as the chain L 3 This 1 of its order --------- line-printed by the Consumers' Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs, the paper size is applicable to China National Standard (CNS) A4 (210 X 297 mm) -12-rhyme 4] 6960 _S! '_ VI. Application scope 12 3 4 12345678901234567890123 45678901234 567890123456789 * · · Two:' · RVLa DIQMTQSPSSLSASVGDRVTITC RASEIIYSYLA WYQQKPGKAPKLLIY FR1 CDR1 t * FR2 5 6 7 8 9 (Please read the note on the back first? page) Printed by 0123456 78901234567890123456789012345678 901234567; 'I' RVLa NAKTLAD GVPSRFSGSGSGTDFTFTISSLQPBDIATYYC QHHFCFPRT CDR2 FR3 CDR3 10. 8901234567- RVLa FGQGTKVEIK FR4 'China's national standard-paper-based (applicable to China) Specifications (210 X 297 mm) -13-Γ 416960 A8 BS C8 'D8 VI. Application for profit scope 1 4. An anti-human IL-8 reconstituted human antibody H chain, which is characterized by containing (1) human antibodies Η chain C region, and (2) a Η chain V region formed by the Η chain CDR of a human monoclonal antibody containing human antibody 8 chain FR ′ and anti-human IL-8, wherein the human antibody Η chain C region Is the human Cr region, the human S-body Η chain FR1 ′ 2, 3 is from the human antibody VDH2 6 ′ human antibody: FR4 is from the human antibody 4B4, and the Η chain CDR has the following amino acid sequence: \ V r CDR 1; Asp Tyr Tyr Leu Ser. CDR 2; Leulie Arj Asn Lys Ala Asn Cly Tyr Thr Arg Glu Tyr Ser Alt Ser Val Lys Gly CDR 3; Glu Asn Tyr Arg Tyr Asp Val Glu Leu'Ala. Tyr 1 5. The reconstituted human antibody according to the scope of the patent application, item 14, wherein the V region of the Η chain is RVHa, RVHb, RVHc * RVH d-RVHe «RVHf» RVHg or RVH h amino acid Sequence or part of it. This paper size applies to China National Standard (CNS) A4 (210 χ 297 male) -nnnn I * 1 ί ί * 1 tr n I n a n »n It nn I Λ I n I n · (Please first Note on this page tftti) G Printed by the Consumer Property Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs A8B8C8D8 1 '41 6960 RVHa ～ h 67890123456789 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs VI. Patent Application 1 2 3 123456789012345678901234567890 12345 EVQLLESGGGLVQPGGSLRLSCAASGFTFS DYYLS. \ FR1 、 CDR1 5 6 012ABC3456789012345 RVHa RVHb RVHc RVHd RVHe RVHf RVHg RVHh WVRQAQGKGLELVG ----------- w „----- p -------- ----- P ----- w— ____ pp ----- W__ ------ PA--W— ----- p ----- w— ----------- W -FR2 LIRNKANGYTRBYSASVKG CDR2 This paper standard is commonly used in SS home standards (CNS &gt; A4 specifications (210 * 297 public love) -15- VI. Patent application scope A8B8SD8 ί · 4 1 6960 100 567890ABC12 t ENYRYDVELAY KVHb _________________________________ : .1— ___________ RVHd -------------------------------- --- -------- RVHe -------------------------------- --------- -RVHf -------------------------------- ----------- RVHg -F- ------------------------------------ RVHh -F ----------- ------------------- ----------- FR3 CDR3 110 '34567890123' 7 8 9 67890123456789012ABC345678901234 RVHa ^ RLTISREDSKNTLYLQMSSLKTEDLAVYYCAR (Read the first and read the back (Items on this page) Order --------- Printed by RVHa ~ h WGQGTLVTVSS FR4 of the Intellectual Property Bureau of the Ministry of Online Economics and Work Cooperatives This paper is applicable to the China Standard for Domestic Standards (CNS) A4 specification &lt; 210x297 %> 6. The scope of patent application is A8 B8 C8 D8 含有 -A 1 6960 containing (A) 8 small L chain (B) -8 6. A reconstituted human antibody against human IL-8, its characteristics Contains (1) the human antibody L chain C region, and (2) the human antibody L chain FR, and an anti-human IL mouse monoclonal antibody 1-chain &lt;: ¥ 11 into a human antibody L chain V region, so: And 'contains (1) the human C chain and k (2) contains the human antibody? R, and the anti-human IL mouse monoclonal antibody Η chain CDR formed by the human Η chain V region 'made Η chain:, the L chain CDR has the amino acid sequence shown below'. CDR 1; Arg Ala , Ser Glu lie lie Tyr Ser Tyr Leu All CDR 2; Asn Ala. Lys Thr leu All Asp-, --- -CDR 3; Gin His His Phe Gly Phe Pro Arg Thr Acid sequence · column: Please read the note J on the back of the book first α; CDR 1 printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs; Asp Tyr Tyr Leu Ser CDR 2; leu lie Arg Asn Lys Ala Asn Gly Tyr Thr Arg Glu Tyr Ser Ala Ser Val Lys Gly CDR 3; Glu Asn Tyr Arg Tyr Asp Val Glu Le.u Ala Tyr17. The reconstructed human antibody according to item 16 of the patent application, wherein the L chain CDR has the amino acid sequence shown below This paper size is applicable to China National Standards (CNS) A4 Regulations (210 X 297 Gong) -17-B8 j 4 t 6960 6. Patent Application Range c DR 1; krg Ala Ser Glu lie lie Tyr Ser Tyr Leu Ala CDR 2; Asn Ala Lys Thr Leu Ala Asp ... CDR3; Gin His His Phe Gly Phe Pro Ar g Thr This H chain CDR has the amino acid sequence shown below: CDR 1; Asp Tyr Tyr Leu Ser v &gt; CD R 2; Leu lie Arg Asn Lys Ala Asn Gly Tyr Thr Arg Glu Tyr Ser Ala Ser Val Lys Gly CDR 3 Glu Asn Tyr κτζ Tyr Asp Val Glu leu Ala Tyr The human antibody L chain FR is from human antibody RE 1; the human antibody Η chain FR1, 2 * 3 is from human antibody VDH26 'FR4 is from human antibody 4B4, the human anti, The C region of the L chain of the human body is the human C λ region: and the C region of the human antibody is the human_C r 1 region * 18. If the reconstituted human antibody of item 16 of the patent application is used, wherein the L chain CDR has the following amine Glycine sequence: CDR 1; Arg All Ser Glu lie lie Tyr Ser Tyr Leu Ala CDR 2; Asn.Ala Lys Thr Leu Ala Asp CDR 3; Gin His His Phe Gly Phe Pro Arg Thr This H chain CDR has the following amino acids Sequence: This paper standard applies to E3S home standard (CNS) A4 specification (210 ^ 297 public love) Please read the precautions on the back before this page) ... / ο Order --------- line Economics; 3-Ministry «? Printed by the Consumer Affairs Cooperative of the Property Bureau -18-I'- 4 1 69 60 A8 B8 C8 D8 VI. Application Scope 1 CDR 1; Asp Tyr Tyr Leu Ser CDR 2; Leu He hrg Asn Lys Ala Asn Gly Tyr Thr Arg Clu Tjrr Ser Ala Ser Yal Lys Gly CDK 3; Giu Asn Tyr Arg Tyr Asp Val Glu Lea AU Tyr The human antibody L The chain FR is derived from the human antibody RE 1; the human antibody Η chain FR1, 2, 3 is derived from the human antibody VDH2 6, and FR4 is derived from the human antibody 4B 4 'The human antibody L chain C region is a human C / C region; and the human The antibody C chain is the human C r 4 region. 19. The reconstructed human antibody 'according to item 16 of the patent application, wherein the L chain V region is an amino acid sequence having RVL a as shown below. (Please read the precautions on the back before filling in this page> 15J.-Line 'Printed by the Intellectual Property Bureau Staff Consumer Cooperatives of the Ministry of Economic Affairs The paper size is applicable to the National Temple of China ^^ Dirty 4 specifications (210 X 297 mm) -19 -μ 4169 69 C8 * r D8 VI.Applicable patent scope 12 3 4 12345678901234567890123 45678901234 567890123456789 RVLa DIQMTQSPSSLSASVGDRVTITC RASEIIYSYLA WYQQKPGKAPKLLIY FR1 'CDR1. &gt; p FR2 5 6 7 8 9 ------------ ii — (Please read the note on the back? Matters before filling out this page) Printed by the Employees 'Cooperatives of the Intellectual Property Bureau of the Ministry of Economy Printed on 0123456 78901234567890123456789012345678 901234567.,' RVLa NAKTLAD GVPSRFSGSGSDFDFTFTISSLQPEDIATYYC QHHFGFPRT CDR2 FR3 CDR3 10; 8901234567- RVIK FG paper for China National Standard (CNS) A4 Specification (210 x 297 mm) Order: Line ·-20-Six A8B8C8D8 f 416960, Patent Application Scope 2 0. Such as the application of the reconstituted human antibody No. 16 of the patent scope 'where the Η chain V Zones have RVHa, RVHb, RVHc, RVHd, RVHe, RVHi, RVHg or The amino acid sequence of RVHh. 1 2, 3 123456789012345678901234567890 12345 Please read the note of Φ RVHa ~ h EVQLLESGGGLVQPGGSLRLSCAASGFTFS DYYLS FR1 CDRI 4 67890123456789 5 6 012ABC3456789012345 RVH RVH RVH RVH RVH RVH RVH RVHg RVHh WVRQAQGKGLELVG LIRNKANGYTREYSASVKG • P- -W- w- One pp ----- W— • P * -W- --W- FR2 CDR2 This paper size applies to Chinese national standards ((: 1 &lt;: 5) 8-4 specifications (210 * 297) &gt; _21_ a | &gt; 416960 C8 ▼ Printed by the Consumers ’Cooperatives of the Ministry of Intellectual Property Bureau ^ I 1 '41 6960 D8 VI. Application for profit-seeking range 2.1 · —A DNA that can encode the anti-human IL-8 antibody chimeric antibody L chain: It is characterized by containing (1) a human antibody [chain 0 region: and (2) Anti-human IL-8 mouse single anti-hip 1 chain ¥ region: made • where the L chain V region is the following amino acid sequence shown in the coding sequence 2 6: Met Ser Val Leu Thr Gin Ya. 1 Lea Gly Leu leu Leu Leu Trp Leu -20 -15 * -10 Thr Gly Ala Ars Cys Asp lie Gin Met Tbr Gin Ser Pro Ala Ser ** 5 -11 5 'i〇Leu Ser Ala Ser Va.1 Giy Glu Thr Val Thr lie Thr Cys Arg Ala 15 20 25 Ser Glu ile lie Tyr Ser Tyr Leu Ala Trp Tyr Gin Gin lys Gin 30 35 _ 40 Gly Lys Ser Pro Gin Leu Leu Val Tyr Asn Ala Lys Thr Leu Ala 45, '50 55 Asp Gly Val Ser Ser Are Phe Ser Gly Ser Gly Ser Gly Thr Gin 60 65 70 Phe Sir Leu Arg lie Ser Ser Leu Gin Pro Glu Asp Fhe Gly Ser 75 80 85 Tyr Tyr Cys Gin His His Phe Gly Phe Pro Ar? Thr Phe Gly Gly. 90 95 100 — Gly Thr Lys Leu Glu Leu Lys 105 (Please pay attention to the notes on the back of Mti before filling this page) a _ · J,-. _! 1 Order. Printing by the staff of the Hydrocarbon Village Smart Finance and Health Bureau Co., Ltd. S a ^ iCMS) A4 specification (210 x 297 male S) _ 23 ~ A8SC8D8 »· i Λ71. 6 9 6 Intellectual property of the Ministry of Economic Affairs The staff of the bureau cancels the cooperative cooperative seal ¾ 6. Apply for the scope of 4 benefits 22. For example, the DNA of the scope of patent application No. 21 *, wherein the L chain V region has the nucleotide sequence shown in sequence 26: ATG AGT GTG CTC ACT CAG GTC. CTG GGC TTG CTG CTG CTG TGG CTT 45 ACA GGT CCC AGA TGT GAC ATC CAG ATG ACT CAG TCT CCA GCC TCC 90 CTA TCT GCA TCT GTG GGA GAA ACT GTC ACC ATC ACA TGT CGA'GCA 135 AGT GAG.ATT ATT TAC AGT TAT TTA GCA TGG TAT CAG CAG AAA CAG 180 \ CGA AAA TCT CCT CAG CTC CTG GTC TAT AAT GCA., UA-ACC TTA GCA 225 GAT GGT CTG TCA TCA AGG ΤΪ0 AGT GGC AGT GGA TCA GGC ACA CAG 270 TTT TCT CTG CGG ATC AGC AGO CTG CAG CCT GAA GAT TTT GGG AGT 315 TAT TAC TGT CAA CAT CAT TTT GGT TTT CCT CGG ACG TTC GGT GGA 360 GGC ACC AAG CTG GAA CTC AAA C __ 382 2 3, an anti-human IL-8 antibody that can be encoded Chimeric antibody H chain DNA: characterized by containing (1) the human antibody 区 chain C region; and (2) anti-human IL-8 mouse monoclonal antibody Η chain V region; made, wherein the Η chain V region is the following amino acid sequence shown in coding sequence 2 7: Met Lys Leu Trp leu Asn Trp Val Phe Leu Vai Thr Leu Leu Asn -19, -15 -10 -5 Gly lie Gin Cys Glu Val Lys Leu Val Glu Ser Gly Gly Gly.Leu -11 5 10 He Gin Pro Gly Asp Ser Leu Arg Leu S-er Cys Yal Thr Ser Gly · 15 20 25 This paper size is applicable to China's 0 Chinese standard (CNS) A4 specification (210 * 297mm S) _ Please read the item ii of the back of the book. I order A8 B8 C8 D8 * 416960 VI. Patent application scope Phe Tlir Phe Ser Asp Tyr Tyr Leu Ser Trp Val krg Gin Pro Pro 30 35 40 (fly Lys Ala Uu Glu Trp Val GLy Leu lie Arg Asn Lys Aia Asn 45 50 55 Gly Tyr Thr Arg GLd Tyr Ser Ser Val Lys Cly krs Phe Thr 6.0. 65 70 τ · He Ser Arg Asp Asp Ser Gin Ser He Leu Tyr Leu Gin Met Asn 75 80, 85 Thr Leu Arg Gly Glu AspSer Ala Thr Tyr ^ Tyr Cys Ala Arg Glu 9095 100 Asd Tyr Arg Tyr Asp Vai.Glu Leu Ala Tyr Trp Gly Gin Gly Thr • 110 115 Leu Val Thr Val Ser Ala 120: 2 4. If applying for a patent The DNA of the range item 23, wherein the H chain V region is a nucleotide sequence having the sequence shown in sequence 27: (Please read first? ? Note on the back page) Printed by ATG AAG TTG GGT ATC CAG ATA CAG CCT TTC ACC TTC GGA AAG.GCA GGT TAC ACA ATC TCC AGA ACC CTG AGA AAC TAT AGO CTG GTC ACT TGG TTA TGT GAG GGG GAT AGT GAT CTT GAG AGA GAG GAT GAT GGT GAG TAC CAC GTC TCT AAC TGG CTG AAA TCT CTG TAC TAC TGG GTG TAC AGT TCC CAA GAC AGT GTA GAG GCA G gtt m CTG GTG AGA CTC CTG AGC 'CCT CTC GCA TCT AGC ATC GCC ACT CTT GCT CH GTG ACA GAG TCT'GGA TCC TGT GTA TGG GTC CGC ΑΠ AGA AAC GTG AAG GGT CTC TAT CH TAT TAC TGT TAC TGG GGC CTT TTA AAT 45 GGA CGC TTG 90 ACC TCT GGG 135 CAG CCT CCA 180 AAA GCC AAT 225 CGG TTC ACC 270 CAA ATG AAC .315 GCA CGA GAG 360 CAA CGG ACT 405 424 Wood paper size applies to China National Standard (CNS) A4 specifications (210 x 297 public love) 25-A8 BS C8 D8 1 6960 Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs • Printing 6. Application for patents 2 5 · — A DNA encoding the L chain V region of a mouse monoclonal antibody against human IL-8, in which the L chain V region Has a sequence The following amino acid sequences are shown in 2 6: Met Ser Val Leu Thr Gin Va. 1 Leu Gly Leu Leu leu Leu Trp Leu -20 -15 -10 Thr Gly Ala Arg Cys Asp lie Gin Met Thr Gin Ser Pro Ala Ser- 5 -1 1 ； \ 5 10 Leu Scr Ala Ser Yal Gly Glu Thr Val Thr &gt; Ile Thr Cys Arg AU 15 20 25 Ser Glu lie lie Tyr Ser Tyr Leu Ala Trp Tyr Gin Gin iys Gin 30 35 40 Gly Lys Ser Pro Gin Leu Leu Val Tyr Asn Ala Lys Thr Leu AU 45 50 55 Asp GJy Val Ser Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Gin 60 65-70 Phe Ser Leu Arg lie Ser Ser Leu Gin Pro Glu Asp Phe Gly Ser 75 80 85 Tyr Tyr Cys Gin His His Phe Gly Phe Pro Arg Thr Phe Gly Gly • _-90 95 100 Gly Thr Lys Leu Glu Leu Lys,: 105 * 26. For example, please request the DMA of the 25th item of the patent scope, which encodes the L chain The DMA of the V region has the following nucleotide sequence shown in sequence 26: ATG AGT GTG CTC-ACT CAG GTC CTC GGG HG CTG CTC CTC TGC CTT 45. ACA GGT GCC AGA TGT GAC ATC CAG ATG ACT CAG TCT .CCA GCC TCC 90 (please read the note on the back first) Alpha line:-This paper standard is in accordance with China Standard for Household Standards (CNS) A4 (210 * 297 male dragons) _ 26 — 6. Application for patent scope A8 B8 C8 DS 416960 CTA TCT GCA TCT CTG GGA GAA ACT GTC ACC ATC ACA TGT CGA GCA 135 AGT GAG ATT ATT TAC AGT TAT TTA GCA · TGG TAT CAG CAG AAA CAG 180 GGA AAA TCT CCT CAG CTC CTG GTC TAT AAT GCA AAA ACC TTA GCA 225 .GAT GGT GTG TCA TCA AGG Π0 ACT GGC ACT CGA TCA GCA ACA CAG 270 '2t ΤΠ TCT CTG CGG ATC' AGO AGC CTG CAG CCT GAA Qkl ΠΤ- GGG AGT 315: TAT TAC TGT CAA CAT CAT TTT GGT ΤΠ CCT CGG ACG TTC GGT GGA 360 GGC ACC AAG CTG GAA CTC AAA C 1 · 382 *-· _ ·. Y — &gt; · 27.—A DNA encoding the H chain V region of a mouse monoclonal antibody against human IL-8, wherein the V region of the Η bond is shown in sequence 2 7 Amino acid sequence: Please read the is. On the back. * Matters then. Scripture-Printed by Met Ly $ Leu Trp Leu Asn Trp Vai Phe Leu Yal Thr Leu Leu Asn -19 -15 -10 -5 Gly He Gin Cys Gla Val Lys Leu 'Val Glu Ser Gly Gly Gly Leu -1 1 5 10 He Gin Pro Gly Asp Ser Leu Arg Leu Ser Cys Val Thr Ser Gly 15 20 25 Phe Thr Phe Ser Asp Tyr Tyr Leu Ser Trp Val Arg Gin Pro Pro 30 35 40 Ciy Lys Ala Leu Clu Trp Val Gly Leu lie Arg Asd Lys Ala Asn 45 . 50 55 Gly Tyr Thr Arg Clu Tyr Ser Ala Ser Val Lys Gly hrg Phe Thr 60 '65 70 lie Ser Arg Asp Asp Ser Gin Ser lie Leu Tyr Leu Gin Met Asn: * 75 80 85 G thread paper size applicable to China Standard (CNS) A4 Specification &lt; 210 * 297 GM &gt; — 27 _ Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs Δp ^ g Γ 416960 D8 ·. 6. Apply for the scope of Bailey Thr Luu Arg Gly GU Asp Ser Ala Thr Tyr Tyr Cys Ala Αγε Glu 90 95 100 Asn Tyr Arg Tyr Asp Val Glu. Leu Ala Tyr Trp Gly CLa Gly Thr • 105: 110 115 Leu Val Thr Val Ser Ala 120 — 28. If the scope of the patent application is No. 27 DNA, wherein the DNA encoding the Η chain V region has the following nucleotide sequence shown in sequence 2 7 ATG AAG TTG TGG TTA AAC TGG GTT TTT TZG GTA ACA CTT TTA AAT 45 GGT ATC CAG TGT GAG GTG AAA CTG GTG GAG TCT GGA GGA GGC TTG 90 ΑΤΑ CAG CCT GGG GAT TCT CTG AGA CTC TCC TGT GTA ACC. TCT GGG 135 TTC ACC TTC AGT GAT TAC TAC TAC CTG AGC TGG GTC CGC CAG CCT CCA 180 GGA AAG GCA CTT GAG TGG GTG GGT CTC m AGA AAC AAA GCC AAT 225 GGT TAG ACA AGA GAG TAC AGT GCA TCT GTG AAG GGT CGG TTC ACC 270 ATC TCC AGA GAT GAT TCC CAA AGC ATC CTC TAT CTT CAA ATG AAC 315 ACC CTG AGA GGT GAG GAC AGT GCC ACT TAT TAC TGT GCA CGA GAG 360 AAC TAT AGG TAC GAC GTA GAG CTT GCT TAC TGG GGC CAA GGG ACT 405 CTG GTC ACT GTC TCT GCA G 424 2, 9... 'Can encode anti-human I] L-8 small employment • single antibody CDR DNA of the LV region, of which The CDR can encode the following amine paper sizes applicable to the national standard (CNS) A4 specification (210 x 297 mm) _ 28-!! '1! · Λνι ---- 丨 order .----- --- Γ &lt; Please read the precautions on the back of the page before using this page) A8B8c8n8 4 1 6960 VI. Applying for a profit-oriented acid sequence: CDR 1; Are Ala Ser Glu He. Lie Tyr Ser Tyr -Leu Ala CDR 2 i Asu Ala Lys Thr. Leu Ala Asp CDR 3; Gin His His Phe Gly Phe Pro Arg Thr 30. Please enumerate the DMA of the CDR which can be encoded according to item 29 of the patent, wherein the CDR is a nucleotide sequence having the following sequence:. \-&Gt; 'CDR 1; CGA GCA AGT GAG ATT ATT TAC AGT TAT TTA GCA CDR 2; AAT -GCA AAA ACC TTA GCA GAT • * CDR 3; CAA CAT CAT TTT GGT TTT CCT OGG ACG 3 1. —A DNA that encodes the CDRs of the H chain V region of the anti-human IL-8 mouse monoclonal antibody, where CDR is Can encode the amino acid sequences shown below: 1 C'DR 1; Asp Tyr Tyr Leu Ser. CDR 2; Leu lie Arg Asn Lys Ala Asq Cly Tyr Thr Arg Glu Tyr Ser Ala Ser Vil Lys Gly ·. = CDR 3 Glu Asn Tyr Arg Tyr Asp Val Glu Leu Ala Tyr 3 2. For example, the C.DR-encoding DNA can be applied to the scope of the patent application No. 31, where the CDR has the nucleotide sequence shown below: This paper is applicable to China S House Standard (CNS) A4 Specification (210 «297 mm; 3) _29- — — — — — — — — — — I] V. ^ ————— — II — — —;-(Please first (Notes on the back of the page are read again on page 0) Printed by the Intellectual Property Bureau of the Ministry of Economic Affairs Consumer Cooperatives Printed by the Intellectual Property Bureau of the Ministry of Economic Affairs Printed by Industrial and Commercial Cooperatives § 88 1 4 1 6960 C8 ^ D8 VI. Patent application scope CDR 1; GAT TAG TAG CTG ACC CDR 2; CTC ΑΠ AGA AAC AAA GCC AAT GCT TAG ACA AGA GAG TAC AGT GCA TCT GTG AAG GGT CDR 3; GAG AAC TAT AGG TAC GAC GTA GAG CTT GCT TAG 33. A type of DNA encoding the reconstructed human antibody L chain V region of an anti-human IL-8 antibody, which is characterized by containing (1) the human antibody L chain V region FR and (2) the CDR 'of the L chain V region of the anti-human IL-8 mouse monoclonal antibody, wherein the above CDR is an amino acid sequence having the following: CDR 1; Arg Ala Ser Giu lie lie Tyr Ser Tyr Leu Ala CDR 2; Asn Ala Lys Thr Leu Ala Asp CDR 3 5 Gin His His Ph'e Gly Phe Pro Arg Thr. And the FR is derived from the human antibody REi. 34. The DNA of item 33 in the scope of the patent application, wherein the L chain V region is an amino acid sequence having rvla as shown below. This paper size applies to China National Standard (CNS) A4 specifications (21〇X 2_ ^) &lt; Please read the notes on the back before filling this page); Line-30-Coco's 416960 C8 D8 34 (matters please read the back of the note and then fill in this page) 12345678901234567890123 45678901234 567890123456789 RVLa DIQMTQSPSSLSASVGDRYTiTC RASE1IYSYLA WYQQKPGKAPKLLIY FR1 CDR1 .V · FR2 5 6 7 8 9 0123456 78901234567890123456789012345678 901234567, *, RVLa NAKTLAD GVPSRFSGSGSGTDFTFTISSLQPBDIATYYC QHHFGFPRT CDR2 FR3 CDR3 &gt..; _ 10 8901234567- Printed by RVLa FGQGTKYBIK FR4 by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs This paper is applicable to Chinese National Standard (CNS) A4 (210 X 297 public love) -31--D8! '41 6960 VI. Application 4 Scope 3 5. As the DMA in the scope of the patent application No. 33 3 * It has the following nucleotide sequence shown in Sequence 6 2 or Sequence 6 5: (Please note on the back of Μΐί before this page) (Serial number ： 6 2) ATG GGA TGG ACC TGT ATC ATC CTC- TTC TIQ GTA GCA ACA GOT ACA 45 GGT GTC CAC TCC GAC ATC CAG ATG ACC CAG AGC CCA AGC A &amp; C CTG 90 ACC .GCC AGC GTA GGT GAC AGA GTG ACC ATC ACC TGT CGA GCA AGC- 135 GAG ATT ATT TAG ACT TAT TTA GCA TGG TAC.CAG CAG AAG CCA GGA 180 AA &amp; GCT CCA AAG CTG CTG ATC TAG AAT GCA AM'ACC TTA GCA GAT 225 GGi. GTG CCA AGC AGA TTC AGC GGT AGC GGT AGC GGT ACC GAC TTC 270 ACC TTC ACC ATC AGC AGC CTC CAG CCA GAG CAC ATC GCT ACC TAC 315 TAC TGC CAA CAT CAT TIT GGT TTT CCT CGG ACG TTC GGC CAA GGG 360 ACC AAG GTC GAA ATC AAA C 379 (serial number: 6 5) ATG GGA TGG AGC TGT ATC ATC CTC TTC HG GTA GCA ACA GCT ACA 45 • «CGT GTC CAC TCC GAC ATC CAG ATG ACC CAG AGC CCA AGC AGC CTG 90. I AGC GCC AGC GTA GGT CAC AGA CTG ACC ATC ACC TGT CGA GCA ACC 135 (GAG ATT ATT TAC AGT TAT TTA GCA TGG TAC CAG CAG AAG CCA GCA 180 Printed by co-operatives of the Intellectual Property Bureau of the Ministry of Economy AAG GCT CCA AAG CTG CTG ATC TAC AAT GCA AAA ACC ΠΑ GCA GAT 225 GGA GTG CCA AGC AGA TTC AGC GCT AGC GGT AGC GGT ACC GAC TAC 270 ACC TTC ACC ATC -AGO AGC CTC CAG CCA GAG GAC ATC GCT- ACC TAC 315 TAC TGC CAA CAT 'CAT ΤΠ GGT ITT CCT CGG ACG HC GGC CAA GGG 360 ACC AAG GTC GAA HTC AAA C .379, 3.6-a kind of editable brick anti-human IL- 8 Antibodies Reconstructed DNA of the human antibody Η chain V region, which is characterized by containing. -32-This paper is in the standard of China National Standard (CNS) A4 (210 * 297 mm) A8SC8D8 4 1 6960 6. Application The scope of the patent is (1) the FR of the human antibody Η chain V region, and (2) the anti-human IL-8 mouse monoclonal antibody 所 chain V region of the CD R. The above CDR has the amino acid sequence shown below :. Two CDR 1; Asp Tyr Tyr Leu Ser CDR 2; Leu lie krz Asn Lys Ala Asn Gly Tyr Thr 'Arg Glu Tyr Ser Ala Ser Yal Lys Gly'. \ 'CDR 3; Glu Asn Tyr Arj Tyr Asp VaJ C- Iu Lee Ala Tyr where FR1, 2, 3 are derived from human antibody VDH26, and FR4 is derived from human antibody 4B4. 37. The DNA encoding the reconstructed human antibody Η chain V region according to item 36 of the patent application scope, wherein the Η chain V region may encode RVHa, RVHb · RV'Hc, RVHd, RVHe, RVH f, the amino acid sequence of RVHg or RVHh. (Please note on the back of M1 * first page) Order --------- line 'Printed by the Consumers' Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs Standards of this paper A 0 standard (CNS) A4 specification (210 x 297 public love) „„ -do VI. Application Patent range RVHa ~ h A8SC8D8 6 9 6 i— 4 1 2 123456789012345678901234567890 12345 EVQLLESGGGLVQPGGSLRLSCAASGFTFS DYYLS FR1 CDR1 4 67890123456789 5 012ABC3456789012345 Order-Set up the Ministry of Wisdom. ¾ The staff of the Property Bureau will cooperate to print RVHa RVHb RVHc RVHd RYHe RVHf RVHg RVHh WVRQAQGKGLELVG ----------- W- ----- p- ------- ----- p ----- w— one_ — — one one — one W one one W------ P ----- W--- --------- w__ FR2 LIRNKANGYTREYSASVKG CDR2 4 'This paper size is applicable. 0 National Standard (CNS) A4 size (210x297 male S) _ 34 _ Application for patent scope RVHa RVHb RVHc RVHd RVHe RVHf RVHg RVHh A8 B8 C8 D8 \ ^ 'A \ 696 r \ 7 8 9 67890123456789012ABC345678901234 100 567890ABC12 rltisredskntlylqmsslktedlaVyycar enyrydvelay -F- -F- FR3 CDR3 Please read the details on the back first? (The matter is further wide / this page) · 110 34567890123 Printed by the Consumer Cooperatives of the Intellectual Property Office of the Ministry of Economic Affairs 〆 RVfTa ~ h WGQGTLVTVSS FR4 This paper is in accordance with the China Standard Standard (CNS) A4 (210x297) -35- 696α k C8 D8 · Sixth, the scope of patent application 3 8. If the DN person applying for the scope of patent application No. 36, it is a person with the sequence 38, 41, 44, 45, 48, 51, 54 or 55 (please read the back first Please note that the following nucleotide sequence is shown in this page: "Two (sequence number: 3 8) ATC GAG TTT GGG CTG AGC TGG GTT TTC CH CTT GCT ATT TTA AAG 45 ... GGT CTC CAG TGT GAA GTG CAG CTG TTG GAG TCT GGG GCA GGC TTG 90 GTC CAG CCT GGG GGT TCT CTG AGA CTC TCA TGT GCT CCC TCT GGA 135 TTC ACC TTC Κ0Ί GAT TAC TAC CTG AGC TGG GTC CGC CAA GCT CAA 180 GGG AAA GGG CTA GAG TTG GTC ATT AGA AAC AAA · CCC AAT 225 GGT TAC ACA AGA CAG TAC AGT GCA TCT GTG AAG GGC-AGA CTT ACC 270 ATC TCA AGA GAA GAT TCA AAG AAC ACG CTG TAT CTG CAA ATG AGC .315 AGC CTG A / l \ kQZ GAA CAC TTG GCC GTG TAT TAC TGT CCT AGA GAG 360 AAC TAT CGC TAC GAC GTA GAG CTT GCT TAC TGG GGC CAG CCA ACC 405. CTG GTC ACC GTC TCC TCA G '424 I &lt; (Serial number: 4 1) ATG GAG TTT GGG CTG AGC TGG GTT TTC CTT GTT GCT ATT TTA AAG 45 t Economy Members of the Ministry of Wisdom and Time Bureau 1. Save fees ^ i. GCT GTC CAG TCT GAA GTG CAG CTG TIQ CAG TCT GGG GGA GCC TTG 90 GTC CAG CCT GGG GGT TCT CTC AGA CTC TCA TCT GCT GCC TCT GGA 135 TTC ACC TTC AGT GAT TAC TAC CTG AGC TGG GTC CQC CAA GCT CAA 180 GGG AAA GGG CTA GAG TGG CTG GGT CTC ATT AGA AAC AAA CCC AAT 225 GGT TAC ACA AGA GAG TAC AGT CCA TCT GTG AAG GCC AGA CTT ACC 270 ATC TCA ACA GAA TAT AAC ACG CTG TAT CTG CAA ATC ACC 315 AGC CTG AAA ACC CAA GAC TTG GCC CTG TAT TAC TGT GCT AGA GAG 360 AAC TAT CGC TAC GAC GTA GAG CTT CCT TAC TGG GGC CAG CCA ACC. 405 CTG GTC ACC GTC TCC TCA G 424 This paper is applicable to Θ S Standard (CNS) A4 specifications (210 x 297) & _ 36 Printed jaws by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 416960 C8 D8 VI. Application 4 Benefits (Serial Number: 4 4 ) ATG GAG TTT GGG CTG AGC TGG CTT ΤΪCH GTT GCT ATT ΠΑ AAG 45 GGT GTC CAG TGT GAA GTG CAG CTG π〇GAG TCT GCG GCA GCA TGC 90 GTC CAG CCT GGG CGT TCT CTG AGA CTC TCA TCT GCT GCC TCT GCT TCT GCT TTC ACC TTC AGT GAT TAC TAC CTG AGC TGG GTC CGC CAA GCT CCA 180 GGG AAA GCG CTA GAG TTG GTG CGT CTC ATT AGA AAC AAA GCC AAT 225-CGT TAC ACA AGA GAG TAC AGT GCA TCT GTG AAG GGC AGA CH ACC 270 ATC TCA AGA GAA GAT TCA AAG AAC ACC CTG TAT CTG CAA ATG AGC 315 AGC CTG AAA ACC GAA GAC TTG GCC GTG TAT TAG TGT GCT AGA GAG '360 AAC TAT CGC TAC GAC GTA GAG CTT GCT TAC TGG GGC CAG GGA ACC 405 CTG CTC ACC GTC TCA G 424 (serial number: 4 5) ATC GAG TTT GGG CTG AGC TGG GTT TTC CTT GTT GCT ATT TTA AAG 45 'I GGT GTC CAG TGT CAA GTG CAG CTG TTG GAG TCT GCG GGA CGC HG 90 GTC CAG CCT GGC GGT TCT CTG AGA CTC TCA TCT GCT GCC TCT GGA 135 TTC ACC TTC AGT GAT TAC TAC CTG AGC TGG GTC CGC CAA GCT CCA 180 GGG AAA GGG CTA GAC TGG GTG GGT CTC ATT AGA &quot; AAC AAA GCC AAT 225 CGT TAC ACA AGA TGA GCA TCT GTG AAG GCC AGA CTT ACC · 270 ATC TCA AGA GAA GAT TCA AAG AAC ACG CTG TAT CTG CAA ATG AGC 315 AGC CTG AAA ACC GAA GAC TTG GCC GTG TAT TAC TGT GCT AGA GAG 360! AAC TAT CGC TAC G ^ C CTA GAG CTT GCT TAC TGG GGC CAG GGA ACC 405 CTG CrTC ACC.GTC TCC TCA G 424 This paper size applies to China National Standard (CNS) A4 (210 * 297mm S) _ 37 1 A8B8C8D8 • τ JH6960 VI. Application for 4 benefits (serial number 4 8) ΔΤ &amp; GAG TTT CGG CTG AGC TGG GTT TTC CTT GTT GCT ΑΠ ΠΑ AAC 45 GGT GTC CAG TGT GAA .GTG CAC -CTG.TTG GAG TCT GGti GGA GGC TTG 90 CTC CAG CCT GGG GOT TCT CTG AGA CJC TCA TGT GCT GCC TCT GGA 135 TTC ACC TTC ACT GAT TAC TAC CTG AGC TGG GTC CGC CAA CCT CCA 180 GGG AAA GGG CTA GAG TGG GTG GGT CTC ATT AGA AAC AAA AAT 225 * GOT TAC ACA AGA GAG TAC AGT GCA TCT GTG AAG 'GGC AGA CTT ACC 270 \. ATC TCA AGA GM GAT TCA AAG AAC ACG CTG TAT CTG CAA ATG AGC 315 AGC CTG AAA ACC GAA GAC TTG CCC GTG TAT TAC TGT GCT AGA GAG 360 AAC TAT CGC TAC GAC GTA CAG 0Π GCT TAC TGG CGC CAG GCA ACC 405 CTG QTC ACC GTC TCC TCA G '424 (serial number: 5 1) ATG GAG TTT GGG CTG AGC TGG GTT TTC CTT GTT GCT ATT TTA AAG 45 GGT CTC CAG TGT GAA CTG CAG CTG TTG GAG TCT GGG GGA GGC TTG 90 GTC CAG CCT GGG GGT TCT CTG AGA CTC TCA TGT GCT GCC TCT GGA 135 TTC ACC TTC AGT GAT TAC TAC CTG AGC TGG CTC CGC CAA- GCT CCA 180 CGG AAA CCG CT A GAG TGG CTC GGT CTC ATT AGA AAC AAA GCC AAT. 225 GGT TAC ACA AGA GAG TAC AGT GCA TCT GTG AAG GGC AGA CTT ACC .270 ATC TCA AGA GAA GAT TCA AAG AAC ACG GTG TAT CTC CAA ATC AGC 315 ACC CTG AAA ACC GAA GAC TTC GCC GTG TAT TAC TGT GCT AGA GAG 360 AAC TAT CGC TAC GAC CTA GAG CTT GCT TAC TiiG GGC CAG GGA ACC 405 CTG ^ TC AC £ GJC TCC TCA G 424 Winter Paper &amp; Degree Applicable to Chinese National Standards &lt; 〇 ^) 8 specifications (210 &gt; = 297 meals) _38- &lt; Please refer to the note on the back of * is this page first) tl. ·; 鯓 · Printed by the Intellectual Property Bureau of the Ministry of Economic Affairs Consumer Cooperative Αδ Β8 C8 D8 416960 VI. Application scope (serial number: 5 4) ATG GAG GCT GTC GTC GAG TTC- ACC GGG AAA GGT TAC ATC TCA AGC CTG AAC TAT CTG JJTC TTT GGG CAG TGT CCT GGG TTC AGT CGG CTA ACA AGA AGA GAA AAA ACC CGC TAC ACC ^ TC CTG ACC TGG GAA GTG CAG GGT TCT CTG GAt TAC TAC GAG TGG GTG GAG TAC AGT GAT TCA AAG GAA GAC TTG GAC GTA GAG TCC TCA G GTT TTC CTG TTG AGA CTC CTG AGC GGT CTC GCA TCT AAC ACG CTC CTT GCT CH GTT GCT ATT TTA AAG 45 GA G TCT GGG GGA CGC TTG 90 TCA TCT GCT GCC TCT GGA 135 TGG GTC CGC CAA GCT CCA 180 ATT AGA AAC AAA GCC AAT 225 GTG AAG'GGC AGA TTT ACC 270 CTG TAT CTG CAA ATC AGC 315. &Gt; »TAT TAC TGT GCT AGA CAG 360 TAC TGG GGC CAG GGA ACC 405 424 &lt; Please read the notes on the back before this page)-嗦 · tl: (Serial Number: 5 5 Printed by ATG CAG GGT GTC, Employee Consumer Cooperatives, Intellectual Property Bureau, Ministry of Economic Affairs GTC CAG TTC: ACC GGG AAA CGI: TAC ATC: TCA AGC CTG AAC TAT GTC; GTC TTT GGG CAG TGT CCT GGC TTC AGT GGG CTA ACA AGA AGA GAA AAA ACC CGC TAC ACC CTC CTG .ACC TGG GAA GTG CAG GGT TCT CTG GAT TAC TAC GAG.TGG GTG GAG TAC AGT GAT TCA AAG GAA GAC TTC GAC GTA GAG TCC TCA G GTT TTC CTT GTT GCT CTG TTG GAG TCT GGG ACA CTC TCA TGT CCT CTG ACC TGG GTC CGC GCT CTC ATT AGA AAC GCA TCT AAG GGC AAC ACG CTG TAT CTG CCC GTG TAT TAC TCT CTT GCT TAC TGG GGC ATT ΠΑ AAG GGA CGC HC GCC TCT GGA CAA GCT CAA AAA GCC AAT AGA TTT ACC CAA ATC AGC GCT AGA GAG CAG GGA ACC 45; 90 135 180 225 270 315 360 405 424 -Pound AS B8 C8 m 416960 VI. Patent application scope 39.-A type of DNA encoding the reconstructed human antibody L chain of anti-human i L-8 antibody, which contains (1) the human antibody L chain C region: And (2) the L chain C region formed by the CDRs of human monoclonal antibodies containing human FR and anti-human IL-8; wherein the L chain V region can encode the amino acid of RV La as shown below sequence. _ Please read the back &gt; ί Italian. Page order. Thread. Printed by the Intellectual Property Bureau of the Ministry of Economic Affairs, Consumer Cooperatives. The paper size is applicable to the Chinese National Standard (CNS) A4 specification Γ210 X 297 mm. -40-r 416960 A8 B8 C8 D8 6. Application scope 12 3 4 12345678901234567890123, 45678901234 567890123456789 \ RVLa DIQMTQSPSSLSASVGDRVTITC RASEIIYSYLA WYQQKPGKAPKLLIY FR1 CDR1 FR2 5 6 7 8 9 0123456 78901234567890123456789012345678 901234567 RVLa NAKTLAD GVPSRFSGSGSGTDFTFTISSLQPEDIATYYC QHHPGFPRT CDR2 FR3 CDR3 10 8901234567- Ministry of economic Affairs intellectual property Office employees consumer cooperatives printed RVLa FGQGTKVEIK ,. • · - FR4 this paper The scale is applicable to the China National Standard (CNS) A4 specification (210 x 297 mm) -41 A8BSC8D8 ^ 416960 6. Application for profit-seeking scope 40. For example, the DNA of the 39th scope of the patent application, where the L chain C region has a sequence Nucleic acid sequence shown in 6 2 or sequence 6 5: (sequence number: 6 2) ATG CGA TGG AGC TGT ATC ATC CTC TTC TTG GTA GCA ~ ACA CCT ACA 45 GGT GTC CAC TCC GAC ATC CAG ATG ACC CAG AGC CCA AGC AGC CTG 90 AGC GCC AGC GTA GGT GAC AGA G TG ACC ATC ACC TGT CGA CCA AGC 135 GAG ATT ΑΠ TAC AGT TAT TTA GCA TGG TAC CAG CAG AAG CCA CCA ISO AAG GCT CCA AAG CTC CTG ATC TAC AAT GCA AAA ACC TTA GCA GAT 225 GGA GTG CCA AGC AGA TTC AGC GGT AGC GGT ACC GAC TTC 270 ACC TTC ACC ATC AGC AGC CTC CAG CCA GAG CAC ATC GCT ACC TAC 315 TAC TCC CAA CAT CAT TTT GGT ΤΠ CCT CCG ACG TTC GGC CAA GGG 360. ♦ ACC AAC.GTC CAA ATC AAA C ' 379 (Please read the note on the back. 1) 1 tl: Code sequence / V 5 •• Sr Printed by ATC CGA TGG AGC TGT ATC ATC CTC TCZ TTG GTA GCA KZK GCT ACA 45 CGT GTC CAC TCC GAG ATC CAG ATC ACC CAG AGC CCA AGC AGC CTG 90 AGC GCC ACC GTA GGT GAC AOA GTG ACC ATC ACC TGT CGA CCA AGC 135 GA [: ATT ATT TAC AGT. TAT TTA GCA TGG TAC CAG CAG AAG CCA GGA 180 AAC GCT CCA AAG CTC CTG ATC TAC AAT GCA AAA ACC TTA- GCA GAT 225 GGA GTC CCA AGC AGA TTC AGC GGT AGC GGT AGC GGT ACC CAC TAC 270 ACC TTC ACC ATC AGC AGC CTC CAG CCA GAG GAC ATC CCT ACC TAC 31.5. Storm TAC TGC CAA CAT CAT TTT TTT CCT CCC ACG TTC GGC CAA GGG 360 ACC AAG GTC GAA ATC AAA C 3T9 This paper size applies to China National Standard (CNS) A4 specifications (210 X 297 public love) 42-§; 418960 D8 VI. Application for drought profit range 4 1 The DNA of item 39 or 40 of the patent application 圔 wherein the human antibody L chain C region is a human antibody L chain C / c region. 42. A type of DNA encoding a reconstituted human antibody chain of an anti-human IL-8 antibody, which comprises: (1) a human chain C 「" and (2) a human FR and an anti-human IL-8 Η chain formed by CDRs of a mouse monoclonal antibody \ V region: \ formed, wherein the Η chain V region can encode the following Jin Hu's RVH a, RVHb-RVHc &gt; RVHd '· RVHe * RVHf -RVHg or RVHh amine (Please read the phonetic notes on the back ^ i — I on this page} Printed by the staff of the Intellectual Property Bureau of the Ministry of Economic Affairs and printed by the cooperatives h This paper size applies to China National Standard (CNS) A4 (210x 297 male cages) f 4t6960 VI.Applicable patent range 123456789012345678901234567890 3 12345 RVHa ~ h BVQLLESGGGLVQPGGSLRLSCAASGFTFS DYYLS FR1 'CDR1 4 67890123456789 5 6 012ABC3456789012345 (please read the precautions on the back page first) Consumption cooperation between employees of the Intellectual Property Bureau of the Ministry of Economic Affairs 钍 Printed RVHa RVHb RVHc RVHd RVHe RVHf RVHg RVHh WVRQAQGKGLELVG _____p_ ——W-- ___ One One_ One pp —-— ---- ---------- -W-- FR2 LIRNKANGYTREYSASVKG CDR2 Order · 1111 3 · This paper size is in accordance with China ’s Θ Family Standard (CNS) A4 specification (210x297) -44-VI. Application for patent scope RVHa RVHb RVHc RVHd RVHe RVHf RVHg RVHh gs158 416960 7 8 9 67890123456789012ABC34567890J234 100 567890ABC12 RLTISREDSKNTLYLQMSSLKTEDLAVTYCAR. &Gt; —p — — — — — — _-e _ _____ -F --------------------------- --- FR3 • 110 '34567890123' 'ENYRYDVELAY CDR3 Intellectual Property Bureau, Ministry of Economic Affairs, Industrial and Consumer Cooperation: d'u RVifa. ~ H ffGQGTLVTVSS FR4 This paper has a universal Yin national standard (CNS) A4 specification (2i0x297 gigabytes) AS BS C8 D8 416960 VI. Application for the scope of benefits 4 3. Such as the application of the scope of patent No. 4 2 0 ^^ 厶 ' The 11-strand V region has the following nucleotide sequence shown in the sequence 38, 41, 44, 45, 48, 51 5 4 or 5 5: (sequence number: 3 8) ATG GAG TTT GGG CTG AGC TGG GTT HC CTT GH CCT ATT TTA AAG 45 GCT GTC CAG TCT GAA GTC CAG CTG TTG GAG TCT CGG GGA CGC TTG 90 GTC CAG CCT GGG GGT TCT CTG AGA CTC TCA TGT'.GCT GCC TCT GGA 135 \ TTC ACC TTC AGT GAT TAC TAC CTG AGC TGG GTO CGC CAA CCT CAA '180 -GGG AAA GGG CIA GAG TTG GTG GDI CTC ΑΠ AGA AAC AAA CCC AAT 225, GCT TAC ACA AGA CAQ TAC ACT CCA TCT GTG AAG GGC AGA CTT ACC 270 ATC TCA AGA GAA GAT TCA AAG AAC ACG CTG TAT CTG CAA ATG ACC 315 AQC CTG m ACC GAA CAC TTG CCC CTG TAT TAC TQT GCT AGA GAG 360 AAC TAT CGC TAC CAC GTA GAG CTT GCT TAC TGG GGC CAG- GCA ACC 405 CT.G CTC ACC GJC TCC TCA G 424 ( Serial number: 4 1) 'ATG GAG ΤΤΓ CGG CTC AGC TGG GTT TTC CTT GTT GCT ATT TTA AA G 45 GCT GTC CAC TGT GAA GTG CAG CTG TTG GAG TCT GGG GGA GGC TTG 90. GTC CAG CCT GGG GGT TCT CTG AGA CTC TCA TGT CCT GCC TCT GGA 135 TTC ACC TTC AGT GAT TAC TAC CTG AGC TGG GTC CGC CAA GCT CAA 180 I GGG AAA GGG CTA CAG TCG GTG CGT CTC ATT AGA AAC AAA GCC AAT 225 • GGT TAC ACA AGA CAC TAC ACT GCA TCT CTG AAG GGC AGA CTT ACC 270 ATC TCA AGA GAA GAT TCA AAG AAC ACG CTO TAT CTC CAA ATG ACC 315 AGC CTC AAA ACC GAA CAC TTC GCC GTG TAT TAC TGT CCT AGA CAG 360 AAC TAT CGC TAC GAC CTA CAG CTl GCT TAC TGG GGC CAG OGA ACC 405 CTG GTC ACC GT〇TCC TCA G 424 Common national standard (CNS) A4 Specification (210 * 297) _ 46-(Please read the note on the back ^ Item H Γ 本 I) Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 416960 A8 BS C8 DS Number: 4 4) ATG GAG TTT GGG CTG AGO TGG GTT HC 'CTT GTT CCT ATT TTA AAG. 45 GGT CTC CAG TGT GAA CTG CAG CTC TTG GAG TOT GGC GCA CGC HG 90 CTC CAC CCT GGG CGT TCT CTG AGA -CTC TCA TGT GCT GCC TCT GGA 135 TTC ACC TTC AGT GAT TAC TAC CTG AGC TGG GTC CGC CAA CCT CCA ISO GGG AAA GGG CTA GAG TTG GTG GGT CTC ATT AQA AAC AAA GCC AAT 225 GGT TAC ACA AGA OAG TAC AGT GCA TC Ding GTG AAG GGC AGA CTT ACC 270 ATC AGA GAA GAT TCA AAG AAC ACG CTC TAT CtG CAA ATG AGC 315 AGC CTG AAA ACC CAA GAC TTG GCC GTG TAT TAC TGT GCT ACA DAG 360 AAC TAT CGC TAC GAC CTA GAG CTT GCT TAC TGG CGC CAG GGA ACC 405 CTC GTC GTC TCC TCA G 424 (please note on the back of Mΐί, t matter This page "-#-I * J_. C serial number: 4 5) I ATG GAG TTT GGG CTG AGC TGG GTT TTC CTT GTT GCT ATT TTA AAG 45 GGT GTC CAG TGT CAA GTG CAG CTG TTG CAG TCT GGG GCA CGC TTG 90 GTC CAC CCT GGC GGT TCT CTC AGA CTC TCA TGT GCT GCC- TCT GGA 135 TTC ACC TTC AGT GAT TAC TAC CTG AGC TGG CTC CGC CAA .CCT CCA 180 GGG AAA GGG CTA GAG TGG GTG GGT CTC ΑΠ AGA AAC AAA GCC AAT 225 CGT TAC ACA AGA GAG TAC AGT GCA TGT GTG AAG CGC AGA CTT ACC · 270 ATC TCA AGA CAA GAT TCA AAG AAC ACG CTG TAT CTG CAA ATG AGC 315. AGC CTG AAA ACC GAA GAC TTG GCC GTG TAT TAC TGT CCT AGA CAG 360 AAC TAT CGC TAC GAC GTA GAG CTT GCT TAC TGG GGC CAG GCA ACC 405 CTp GTC AC £ QTC TCC TCA G 424 This paper size applies to China Standard S (CNS) A4 (210 * 297 mm) _ 47 _-鯓-Printed by the Intellectual Property Bureau of the Ministry of Economic Affairs on employee consumer cooperation i 416960 C8 D8, ____ VI. Patent application scope (serial number: 4 8) ATG GAG TTT CGG CTC AGO TGG GTT TTC CTT GTT GCT ATT TTA AAG 45 ' CGT CTC CAG TGT GAA GTG CAG CTG .TTG GAG TCT GG G CGA GGC TTG 90 GTC CAG CCT GCG GCT TCT CTG AGA CTC TCA TCT GCT GCC TCT CGA 135 1TC ACC TTC ACT CAT TAC TAC CTG AGC TGG GTC CGC CAA CCT CCA '180 DG &amp; AAA GGG CTA CAG TGG CTG GCT CTC ΑΠ AAC AAA GCC AAT 225 CGT TAC ACA AGA GAG TAC AGT GCA TCT GTG AAG CGC AGA CTT ACC 270 ATC TCA AGA GAA GAT TCA AACr AAC ACG. CTG TAT CTG CAA ATG AGC '315 AGC CTG AAA ACC GAA GAC TTG GCC.GTG TAC TGT GCT AGA GAG 360 AAC TAT CGC TAC GAC GTA GAG 0Π GCT TAC TGG GGC CAG GGA ACC 405 CTG GTC ACC. GTC TCC TCA G · 424 (Serial number: 5 1) I ATG GAG TTT GGG CTG AGC TGC GTT TTC CTT GTT GCT ATT TTA AAG 45 CGT GTC CAG TCT GAA GTG CAG CTG TTG GAG TCT GGG GGA GGC TTG-90 CTC CAG CCT GCG GGT TCT CTG AGA CTC TCA TGT GCT GCC TCT GGA 135 Member of the Ministry of Economic Affairs Intellectual Property Bureau H Consumer Cooperatives Printed TTC ACC TTC AGT GAT TAC TAC CTG AGC TGG GTC CGC CAA GCT CCA 180 GCC AAA GCG CTA GAG TGG GTC GGT CTC ATT ACA'AAC AAA GCC AAT 225 GGT TAC ACA AGA GAG TAC AGT GCA TCT CTG AAG GGC AGA CTT ACC 270 ATC AGA GAA GAT TCA AAG AAC ACG CTG TAT CTG CAA ATG AGC 315 AGC CTG AAA ACC GAA CAC TTG GCC GTG TAT TAC TGT GCT · AGA GAG 360 AAC TAT CGC TAC GAC CTA GAG CTT GCT TAC TGG CGC CAG GGA ACC 405 CTQ CTC ACC_ GTC TCA G · .424 This paper is commonly used in 0 national standards (CNS) A4 specifications (210x297 public love) _ 48-A8BSC8D8 416S60 6. Application scope (serial number: 5 4) ATC GAG TTT GGG CTG AGC TGO CH TTC CTT GTT GCT ATT TTA AAG 45. GGt CTC CAG TCT GAA CTG CAG CTG HG GAG TCT GGG GGA GGC TTG 90 GTC CAG CCT GOG GCT TCT CTG AGA CTC TCA TGT GCT GCC TCT GGA * 135 TTC ACC TTC ACT GAT TAC TAC CTG AGC TGG GTC CGC CAA GCT CCA 180 GGG AAA GGG CTA GAG TGG GTG GGt CTC ATT AGA AAC kkk GCC AAT 225 GGT TAC ACA AGA GAG TAC AGT GCA TCT GTG AAG GGC AGA TTT ACC 270 ΛΤ0 TCA AGA GAA GAT TCA AAG AAC . CTG CAA ATG AGC 315 AGC CTG AAA ACC GAA GAC TTG GCC GTG 'TAT TAC TGT GCT AGA GAG 360 AAC TAT CGC TAC GAC GTA GAG CTT GCT TAC TGG GGG CAG GGA ACC 405 CT.t CTC ACC GTC TCC TCA G -424 (Serial number: 5 5) \ ATG GAG TTT GGG CTG AGC TGG GTT TTC CTT GTT GCT ATT TTA AAG 45 GGT GTC CAG TGT GAA CTG CAG CTG TTG GAO TCT GGG GGA GGC TTG 90 GTC CAG CCT GGG GGT TCT CTG AGA CTC TCA TGT GCT GCC TCT GGA 135 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs TTC ACC TTC AGT GAT TAC TAC CTC ACC TGG GTC CGC CAA GCT CAA ISO GGG AAA GGC CTA GAG TGG CTC GGT CTC ΑΠ AGA AAC AAA GCC AAT 225 GGT TAC ACA AGA GAG TAC AGT GCA.TCT GTC AAG GGC AGA TTT ACC 270 ATC TCA AGA GAA-GAT TCA AAC AAC ACG CTG TAT CTG CAA ATG AGC 315 AGC CTG AAA ACC GAA GAC TTC CCC GTC TAT TAC TGT GCT AGA GAG 360 AAC TAT CGC TAC GAC GTA GAG CTT GCT-TAC TGG GGC CAG GGA ACC 405 CTG GTC ACC CTC TCC TCA G a〇a -49-This paper standard applies to S gardener standards (CNS &gt; A4 specifications (210 X 297)) Employees of the Intellectual Property Bureau of the Ministry of Economic Affairs Printed by the Consumer Cooperative I 416960 cs D8 6. Scope of patent application 44. For example, the DNA of item 42 or 43 of the patent application scope, wherein the human antibody chain C region is the human antibody chain C 1 region. 4 5. The DNA of claim 42 or 43 in the scope of the patent application, wherein the human antibody Η chain C region is a human antibody C chain C τ 4 region. 4 6. —A method for producing a reconstituted human antibody against human IL-8, which is characterized by including a performance vector containing _DNA as described in any one of claims 39, _4 0 ′ 41 ( HEF-R VL a-g / c), and a expression vector (HEF-RVHa-g r 1, HEF) formed by containing a DNA such as any one of claims 42, 4, 3, 44 or 45 — RVHb-gri -HEF-RVHc-grl * HEF-RVHd-grl, HEF-RVHe ~ grl -HEF-RVHf-grl, HEF-RVHg -gr 1), while transforming COS host cells or CHO host cells, cultured The transformed host cell is obtained by the step of recovering the antibody of interest. 47. A method for producing a reconstituted human antibody against human IL-8, which is characterized by containing DNA such as the scope of patent application No. 39, 40 or 41, and by containing DNA such as the scope of patent application No. 42 A single expression vector formed by DNA of items 4, 3, 4, 4 or 45. A single expression vector is inserted into the single expression vector, the L chain, using at least dhfr as a selectable marker, and the expression vector expressed under the control of the H CMV promoter. At the same time, CΗ0 host cells were transformed, and the transformed host cells were cultured to recover the antibody of interest. 4 8 .-- Manufacturing of reconstituted human antibodies against human IL-8 This paper is applicable to China Standard (CNS) A4 specification (210 X 297 mm) -------- Order --------- line. &Lt; Please read the precautions on the back page first) -50-A8 I f &gt; 41B9B0 VI. Patent Application Scope Method, which is characterized by the simultaneous transformation of CH by a expression vector formed by DNA containing RVL a as in the patent application item 9 and RVHg as in the patent application item 11 ◦ host cell It is formed by culturing the transformed CHO host cell to recover the antibody of interest. 49_ A method for manufacturing a reconstituted human antibody against human IL-8, which is characterized by using an expression vector (HCMV- hWS4 — rla), and transformed into CHO host cells at the same time, and the transformed host cells are cultured to recover the target antibody. Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs This paper is sized to the Chinese National Standard (CNS) A4 (210 X 297 mm) -51-