JPS59500986A - Monoclonal antibody mixture and use of said monoclonal antibody mixture for sensitive immunoassays - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Monoclonal antibody mixture and said monoclonal for sensitive immunoassays Use of a mixture of antibodies es of dealth, United States Department t of Health and Human 5eryices, lcD assistant HD-13496, HD-15454 and CA-26636, which received funding. This is something that was picked up during the research process.

Technical background of the invention Development of hybridoma technology [G, 1 (Ohler and C, mlstein, (1975) Nature 256:495: and J, W, Godi ng, (198 Immunological Methods 39: 2 85], c, p, the only anti) JR' Medicine was provided. The use of these reagents as biochemical intermediates and immunological tools is However, in radioimmunoassay, compared to serum antibodies, Hirohara Affi There were many cases where it was not effective due to low nitei (parental note). (J, W, Goding.

(198 Immuno’logical Methods 39: 285 3゜more hostile]・If you follow the Ibridoma selection procedure, it will be a general VCv monochrome The affinity of the null antibody was the same as above. Furthermore, by producing monoclonal antibodies, The dead can be analyzed by analyzing all the individual responses to I/l1m. It was. [l'lJ, A, 5taines and A, N1. Lew (19 80), Immunology 40: 287] o As a result, 4' Regarding the individual antibodies, the antiserum is the sum of the individual antibodies, I have a good understanding of the CIT index, which can have different attributes. Alof.

Immunochemistry of human ciliary gonadotropin (hcG). A monochrome view of the relative distribution of hormone-receptor groups and changes in hormone-receptor interactions. In the course of systematic research on the effects of monoclonal antibodies, The apparent affinity of the fart partition is compared to the affinity of antibodies on one side! Same as before Hosomi was worried about going up.

Summary of the invention Binding to different antigenic sites of antigens At least two monoclonal anti-monoclonal antibodies of MX#I should be used in a valid assay blood test. The monoclonal antibody fragments that are produced are the same as those of the side effects of the assay against the antigen. It's for sale.

The above-mentioned rapid mixture 1g is sometimes expressed as a gluteal antigen, ν1j is the species of the antigen. Human chorionic gonadotrophs consisting of the amino acid sequences of each antigenic site /) 3 different antigenic sites Polymers such as pins, egg stimulatory hormones, thyroid stimulating hormones or thyroid-forming hormones Useful for assays against peptides. As described above for clinically important antigen assays Using mixtures such as A simple explanation of the Samurai drawing with better measurement sensitivity compared to Figure 1: Radioimmunoas of human chorionic gonadothrobin by two-antibody liquid phase assay. Tusei. In the absence of unlabeled inhibitory factors, labeled hCG bound to Meng 1, antibody or The antibody was killed at approximately 1a] in the valley of the proboscis. This 0111 foot is antibody bath Keimata, - Each mixture was prepared by appropriate dilution. The second antibody is Upugi anti- It happened rarely with mouse I'gG. ED6o plus/for each antibody or mixture Minus 昬 Waseda Soso started. The ED of the interception between BIOI and B102. is 2 ．． 44±0.74X10M, one enemy B101 is 2,88 0.74Xl 0 1VI, 1.058±Q for antibody B102, 995XBIOI, C is antibody B] 5 catchard analysis oMi gradient for 02's hCG +2 (f and half [fii f i d) is A: 5.4X l O’; B : 5. l X 10': C: 1.9 X 1 t)".

Figure 3A: Radioimmunoassay of hCG by two-antibody liquid phase assay. First evil It used the same gonohorobue as the one used in the song. ED, o of Shiriza AlO2 is 8.52±2.94 X 10 rVi, 2.88 ± 0.74 X 10 M for antibody B101, antibody For the mixture of B101 and AlO2, it is 4.66±1.99X10M.

Figure 3B: Antibody B101. B103. Mixture B101+B103’z Inunudai Radioimmunoassay of hCG by two-antibody fL phase assay. The place in Figure 1 The same procedure was used.

m: Radioimmunoassay of hCG using solid eye assay.

A: Antibodies B101 and B102 and mixture 43101+13102°B: Antibody B 102 and B103 and mixture B102+B103゜g5 Figure: Papain digestion B Radioimmunoassay of hCG by two-antibody liquid phase assay using 102i. 1st The same procedure as in Figure '68 was used, except that for precipitating antibody-antigen receptors, Goat anti-mouse F (ab'J2k) was used as the second antibody. There was no effect on the affinity of the mixture even after changing it to sea urchin (results not shown).

Figure 6: B 101-F(ab')2'<h by two-antibody liquid phase assay using Radioimmunoassay of CG. The same hand and shell hormones as in Figure 5) A: A mixture of B101 and B102 and B: Various types of 1&hairy gonadotropin. When mixed with a monoclonal antibody corresponding to the epitope, the measurement of antigen binding assay is It is evident that the same as above for constant sensitivity can be obtained. Depending on the choice of antibody group, there may be some confusion. Affinity of monoclonal antibodies assayed separately It can be made 10 times as large as A.

Affinity segregated in this way can be used in solid-phase assays and liquid-phase two-antibody radioimmune assays. It can also be detected in the assay chair.

The affinity mechanism relies on the formation of multicomponent complexes. combine at the same time Even if two types of antibodies are mixed, affinity does not occur, but Using a combination of antibodies that bind together under appropriate conditions allows for anti-I, KM assays. The measurement sensitivity of B is improved. Combinations of antibodies created by mixing to create affinity If you replace it with the F (aJ fragment) of that antibody, it will no longer be improved. It becomes e. This does not seem to be due to an allosteric effect. I will show you that. The F(ab')zffr fragment forms a specie when mixed with another antibody. However, it does not show the same effect as the complete (1 ntact) antibody.

Based on these detailed examinations and observations, the present invention has developed a t-monoprotein that is useful for highly sensitive assays of antigens. A mixture of clonal antibodies is provided. Antibodies useful for the detection of defects of the present invention may also be denatured. Antibodies that can bind to antigens are prepared under standard conditions that do not cause antigen-binding and in a separate frying tray described below. It is assumed that the antigen can be bound under both conditions under which binding is not possible.

The main text states that two or more types of monoclonal antibodies are used as the TGV rule of Ki and Bo. Ru. The antibodies that are attached to the different antigenic sites of the antigen are 3, 3, 4, 1, 1, 2 and 3 A mixture containing more than n? I is a polypeptide with multiple subunits. It would be useful for loading into assays for large molecules. However, with the following description,... As the fruit of light ν1j, I congratulate you on the mixture mentioned above. .

Seven antibodies of a monoclonal antibody were used in one assay. Although the discovery of this type of antigen was very difficult due to 0J', the present invention A polypep consisting of individual true amino acids of libertide (c) Human chorion 1 is used as a source of this antigen, which is sometimes suitable for the emergence of typhoid antigens. Nadotropin (hCG).

Hormone (TSI (,) and yellow Body formation hormone (LH), which is related to the drowsiness hormone, is 1 bowl shul.

M effective assay τ rows/so that each of the monoclonal antibodies in the assay mixture should be simultaneously attached to enough antibodies, i.e., antibodies, to cause significant binding to the antigen. It should be present in an amount about 10% τ greater than that of the antigen. Preferably, the bond The base of the antigen is higher, for example in the range of about 20 to about 80% or more. Dena σn must be.

In order to obtain the sweat antigen of FiiJ, we first need to develop an antibody against the antigen. Based on the affinity, the net front of the valley antibody can be changed over a wide range, i.e., high affinity. Antibodies that have low affinity have low affinity, and there are also few antibodies that have low affinity. .

The relative size between monoclonal antibodies can also vary over a wide range of timescales. 2 types In the case of a mixture of antibodies of the same type, this value may vary from 106:l to 1:10'. Attend. This is considered to be a strong factor in the decision to custom order the monoclonal antibody. Based on the binding constant, the range is preferably from 102:1 to 1:10" Deriro. In general, the preferred amount of antibody against another antibody is It is probably the same as the ratio of binding constants of antibodies. Therefore, monoclonal antibodies A and When the bond IDs of A and B are 10 and 10, respectively, the relative amount of A and B is approximately 10/ 10 = 10. That is, 1 molecule of A per 8100 molecules.

Detailed description of α subunit and β unit of human ciliary gonadotropin When a polypeptide antigen is raised, each antibody has a different subunit of the antigen. A mixture of monoclonal antibodies can be used to test the VCC site using the mixture. Sei's measurement 4 sensitivity τ Same as above. This is probably due to the appropriate conditions for the antibody. This is because in F, it is possible to use antigens at the same time.

The kelp is used in the assay in its previous form. Stool is a solid matrix It can be used in the form of a face piece with a rough plate, or it can be melted in a suitable back-up solution. It is also possible to use one change in the form of a ``sufu form''.

The presence of antigens in a mixture or sample can be determined quantitatively or quantitatively. For this purpose, conventional immunoassays such as radioimmunoassays can be used . In such a method, the mixture and the antigen can be detected by +-s4. Grease the sample with the mixture in a suitable manner to form a mixture. one specific use As for the development of human gonadotropins for pregnancy tests, .

Fukaimei-i! In order to help Byo, Ichisei's rice grains are shown below.

Is Tensui of course a naughty representative of the uninvented sheep? +J, which completely limits the scope of the present invention. I can't help but interpret it as such. The scope of the present invention is determined by the claims d below. Limited.

Materials and methods Production of monoclonal antibodies: method of vvands and Zurawski (J, L vVands+ V, R, ZurawskL (198 Gastroent erolog)r　80　:　225]　f for Ba1b/c mice intraperitoneal injection of hCG subunits in complete Freund's adjuvant once a month. Immunization was given several times over several months. The tile was given an intravenous booster antigen 3 days before removal. .

(Raw 4 noble salt water 11: +50μ)).

4 rules P 3 t’J Sl/T-A 4-1 Combined with myeloma reduction, Hybridoma cells were isolated using known procedures (J, R, VVandg, V.

R, Zurawski + (1981) Gastroenterology 80: 225: A, tVIarshak-Rothatein%, (1979) J. Immunol. 122: 2491. later childhood Antibodies used in children were purified twice by constant dispersion on Ba1b/c3T3 monolayers. The Hypuridoma axis threading sword and the like are simple and easy to use. All 1 antibody The letters, three numbers, and VC make 'la cup. The letter B indicates that the antibody is β-sub of hCG. Something specific to the unit? The antibody binds to the α-Sag unit in the father and son. Show that.

Cell supernatants or partially purified or purified antibodies or both were used. - minutes 2. In order to attend to the Glass Enemy Antibodies.・L/Go's bloodless feeling of saying M's serum albumin Hybridoma cells were grown in soil, and charcoal with 0.051 Vi of YW was added to water-based water. Dialyzed against ammonium.

偲# Dry gland trowel, desired amount of 0.3M phosphoric acid power Lucimba Sofa, Box 7, 5 CF I protected the powder with 3. In order to obtain purified antibodies, To make violet of rubumin 0.1 rt9/-, use the same protocol, Albumin Total DEAE Affi-Gel Blue Column (Bio-Lad Laboratories).

Two antibodies Radioimmunoas 7Se (: 501ito”’I-hCG and 50 μt unlabeled CG and (both were tested in 1% horse serum and 99% phosphate-buffered saline) 0.3 M calcium phosphate (pH 7.5) of IL) Oμt. Ta. Next, add 100μt of antibody (diluted in 1% horse serum) and remove the tube for 37 hours. Incubate for 1 hour at mouse serum (in phosphorus-salted buffered saline and an appropriate amount of rabbit anti-mouse IgG or The complex was precipitated with goat anti-mouse F (ab')!. Then (immediately) settle the precipitate stubbornly at 37°C for 10 minutes and at room temperature for 1 hour. Roughly count 7ζ.

Facial radioimmunoassay: A plastic microtiter well is coated with an antibody. Prepare a Cooke microtiter plate (with a “U” well Incubate at 5°C for 18 hours and wash the wells 3 times with distilled water. After purifying the excess Izuo antibody, I tasted it. Geras has the potential to cause protein condensation. Although the tech part τ is a trap that completely ranges, the great is 10% d-sou. γ-G-free Wama dish r* (90% phosphoric acid; I tasted it with distilled water. L-gi horse Jf saying bcG of raccoon's t! Lr in progress 50μt of I-hCG was added to the hcG i:i at 5°C. At 8 o'clock [i), the insects were drying, and the radiation of =m+, murmuring J, and saying Great. Fight with distilled water and count the wael/hi.

Sand inch assay: Monoclonal anti-deficiency of at least aotiW/ml Antibody purple It was layered around the plastic surface.

Because it lasted for 4 hours at 37°C, bath sales were booming, and 7 tons of bovine serum albumin. 6-Kr1 50+nM (7℃%JaCte'JE (BSS A-Physiology Award Narusui Moon (Plate TF receiving side, remaining non-standard position on the plastic side ↓ and see the part I took out a light bond. This hormone (50μt J3S A - Physiological S: Add 1 μDa to the microtiter well in the surrounding water for 2 hours. Himetou spoke. This work will be done in 12 minutes, with a round, 5, and 40mm round hUG. The antibody that was layered on the stick was injected into a female. dSA-Wang Li Prize salt water bathing plate I fought against him and took Sosaku's hCG. The body that was marked with a number of symptoms was completely cooled and turned into dsA-4 ! 50,000 to ioo, oo of salt water bath liquid A.

Maintain cpm at room temperature for 2 hours at room temperature, and then plate with BSA-treated 4-fed rice. , the unsweetened labels were removed. Micro titer well All scissors are used for success Then, the total radiation Ke of the plate layer was measured in the valley well.

Alternative materials and methods: Zheji's method [G, M, Edelrn illusion η and J, J .

+vlarcharonis, (1967) A4methods in Imm unology and immunochemistry.

V’o1. I, page 405, Academic Press, view Antibody B102 was digested using YorkJk. Digestive proboscis NaDod 5O4 polyacrylamide gel pneumophoresis (K, vVeber and rA, 0sborn, (1969) J, 8io1.C hem, 244:4406], it is completely 1 (frequently, there is no personnel left at all) It turned out that there was no such thing. Antibody t31.01 F (ab’ h tlT piece) To decorate? This (yo, pepsin digestion CG, Gorini4, (1969) J , Immunol, 103:1132] f was used, however, Vegcin The weight of the antibody was made 2%*% of the weight of the antibody. The remaining L7j antibody was digested with p+(8,1 Protein A-8 epharose (PharmaciaFine Chemicals), Protein A-8ep The aggregate of harose and antibody was removed by centrifugation or VC. Ey temple is enshrined Conditions used CP, L, Ey No. (1978) Immunochemistry 15: 429. ], except that column chromatography A batch process was also used instead. 7I in the presence of a reducing agent d　When applied to S04 polyacrylamide gel, the molecular weight is 105,000. There was only one major band in existence. At 2801m, Gengakuseiho, Amino Kaiping Analysis and V-ko hCG was measured once.

Chloramine-Tf:1ffi, manufactured at Greenwood Temple 11 [F, Greenwood Temple, (1963, l Bioche m, J, 39:114] hCG was iodinated.

fence confusion Radioimmunoassay: Radioactive and cumulative hCG is used as monoclonal antibody B. 01, B102 and a mixture of B101 and B102. The effect of hCG on suppressing the CG was 20 times more active in inhibiting radiolabeled debris. 5c of data If Lfl is mixed in atchard analysis? 1) Compared to l3101 alone, the same The affinity of Ruki is shown, and compared to B102 alone, Ruto is more than 0 years old. (Second figure. Since the affinity was 1, a 1:1 mixture of 21 antibodies was used. Dilute the substance 9 times and bind the same child's tracer with one antibody (11: Achieve 7ri ≦ vomitable. As shown in Table 1, these results are unfortunately not very reproducible. No data shown in the text〃S, Ability of the two raccoon antibodies to exhibit such synergistic effects. is limited to combinations of antibodies that bind to the same long block unit. Does this mean that the combination of AlO2 and B102) has this exciting interaction? It is clear that With a different combination of antibodies, the same feed force was produced 117:)s　fC 0, that is, dlol and Al 02 (Figure 3A 9 father is Bioi and B103 (3B In the same case as in Figure), a similar trend effect was not observed.

Table 1 A mixture of antibodies B101 and B102? Antibodies B101 and B102 compared to I Finity d-needle analysis* Measured using 5 catchard plot [G, 5catchard+ (1949) Anu, N, Y, Acad, Sc i, 51: 660] **KeIHk of mixture of B101 and B102 Ke1 of B101! 1 sai divided by. Same as above? ! - Mixed for the sake of rising Comparing the affinity of a substance with that of an antibody that has the affinity of a delicious sword are doing.

A compound of gioi, AlO2, and norr (the father is a mixture of BltJl and B103) , well, in a phase radioimmunoassay that gave an intermediate line between the two antibodies. However, it is clear that the heather rice is +cT1g (Fig. 4, 9°) 8? It is unlikely that any antibody with lY alone would have high hCG affinity. It was celebrated. However, the migration between antibody 8101 and the mixture was only about 4 times. Antibody B The combination of 102 and B103 did not cause affinity Ruka.

Sandwich assay: Theta Samurai explaining the mechanism of affinity change In order to identify Examples were added (Table n).

If the unlabeled antibody and the radioactive 8154 iJ antibody are injected into the same site, the unlabeled antibody The radioactivity label on the body is 1]. My wife has different antibodies on both sides. Multiple radio labels are tied to the glass VC on the lifting platform that is suspended in the area where it is placed. that the labeled B101 and the non-labeled BIOI are delivered as antigens at the same time; I can't do it. In this case, little radioactive label bound to the plastic is observed. Colleague's outcome ρ also when using n not labeled B102 and non-labeled B102 and t 1 Rinsa nta. , Antibodies B102 and B103 are both dlol and plastic. m　Because I was sweating, I made hcGK sweetfish at a site that is wider than the binding site of BIOI. Change , sign B construction 02. However, it bound to hCG bound to unlabeled B101. in contrast Antibody A102f blocked the binding of radioactive 13101. This means that AlO2 and B 101 and 〃S cannot be applied to hcG at the same time, and once again, antibody B102 It was also noted that Hosomi and B103 could not be concluded at the same time. Encourage island-like interaction For this purpose, it is necessary for the antibody to bind simultaneously at the two hcG sites. However, FJJ said that it was not enough.

The infancy of antibody fragments: monochrome to study the influence of antibody formation on dynamic injection A large number of antibodies were prepared. When digested with antibody 8102t papain, 8101 The ability of antibody B102 to improve affinity was demonstrated (Figure 5). However, by papain digestion of B102, 13102 was isolated to hcG. There was no chivalrous influence on Tei. This is to improve affinity. This shows that the Flc chain acid or double strand of the antibody is required. Bivalent F (ab ’) z to the extent that it has the same affinity as the one above, but the complete antibody e= Ineffective (Figure 6 and Table 1゜Table 1 Combination of monoclonal antibodies-e-2 The number of sandwich assays used was 1. A liquid layer is formed on the surface of the plastic micromechanical plate. cpm of antibody that specifically adhered to the antibody (average value of 3 measurements).Introduced non-typically. Amaishi fcH receptor (i.e., gluing in the presence of a monoclonal antibody that does not bind to hCG) The glue bonded to the plastic did not rub off.

*pku・001: All others are zero and there is no significant slowness.

Surface layer f　5catchard)゛2 identification CG using Ronotowo, 5catchard .

(1949) Ann, N, Y, Acad, Sci, 52: 660 J** B101 divided by Keq of B101 F (ab’)2 and B102 8 eq of a mixture with.

*** KeqQ value of B l 01 F(ab')2 is BIOI Keq in Table I There is no difference between being on shift and being lazy.

Consideration Mixtures of monoclonal antibodies are sensitive to important differences compared to their individual components. Attend on your shoulders. Affection of hybridoma antibody mixture (obviously, a sword). Another load of monoclonal antibodies i), more than one antibody Significant improvements have been made through the use of ridoma antibodies. Hzbe r temple (E, fh ber 4, (198 Resolving antigenjc 5ites and purifying proteins with monoclo nalantibodies”+ +VIonoclonal AnLibod ies in P2ndocrine Re5earch, RavenPre ss] An assay using two monoclonal antibodies and using each antibody separately. Seiniji also launched a series of immunometric tests that were highly controversial.

f (oward temple (J, C, downward%, (1979) Immunol logical Rev, 47: 139 3 is the # of monoclonal anti-salary! i He also showed the endnotes on the bathing screen, proving that he was a servant. Benefits of monoclonal antibodies The ability to react with the base determinant of the one on which the dot is attached may be greatly increased depending on the situation. However, there are many advantages to using FIT-opened anti-plate serum serum.

In addition to the spectacular view of the sea, it is also involved in the anti-seizure conotrol and the in-vivo rabbitfish valley. I would. A mixture of antibodies, Since different quantities are used, another 5 answers are considered to produce a mixed fragment of the antibody "F/l". I'll choose. iH, the cause of affinity R18 on desk m is CE, Kar ush, (1978) Comprehensive Immunology 5 + Immunoglobulins pp, 85 + Plenum tvl edical Book co, fJew York Another antibody present in the dish 7W, but not in the dish 5 stimulated with C and nimeru. Antibody rJ' produced by the same affinity association as above/1. I don't know if it's true. do not have.

A large number of tests were carried out regarding the antibody mixture of Eikai. Antibody B101 and antibody AlO2 ? The combined lift piece, the amount of pre-bu ω, the valley l' line meets t'L, that is, the logic. In the top plot, n is the mixed control line; it exists between f of the BB section of the individual antibodies. Ta. , t3101, AlO2, and mahCG at the same time, I don't even bother V. . During the VC cycle of this hormone, T5 is present with antibodies 13101, B103, and τ Lifting table, logit plot of mixture 1, low resistance yA, low resistance CT engineering affinity resistance The plot follows that of a low-affinity antibody at high yields.

α-sections of antibodies GIUI and 13102 were obtained using a mixture of 35 individual antibodies. Table 1 [Volume As expected, this result will make us pay close attention to the fact that even the VCs who are currently in the process of revising the program are being criticized.

The elucidation of the mechanism is +%1rdAK because the dynamic action g fEb = Usudasa n Become. If F(breath b) laboratory test is used instead of antibody, fco will decrease slightly to the same extent as above. cv) Shows the two layers of vehicle design related to work, engineering, and cooperation.

First, large F'(ab,) engineered antibody fragments are available. Second, one antibody It is believed that the binding of these molecules changes the distribution of hormones and impairs the affinity of the second antibody. T'nozai (allosteric model) is not ot'L. F’ to a different way of thinking L. It is a blessing that the above occurs due to the intermolecular bias between the antibody and the antigen in 21 songs. I'm not worried that I might get mouth disease. (Rie Shio is the molecular force of two antibodies and two antigens. ) is formed, forming a staghorn-like amalgamation. Schumaker temple n-bako I think that the cough is a stable combination of coughs (1111 and n1 [V, N.

Schumaker @, (1973) Immunochewistry 1 0:521]) OF(abtsu2) This suggests that there is no single mechanism for this. Yota. F(c) chain acid phase of the combined shell Although the interaction is consistent with all the data from the phase radioimmunoassay, there are no significant differences. When the assay was completed, it was fixed to the plastic with a plate of 5 pieces and 2 pieces as above. Since this still occurs, we believe that the FIc) interaction is the only safe cause and that 7L. I wonder if the mechanisms are different between the four strips of the two types of assays. do not have. Therefore, the results regarding antibodies dlul and B''103, i.e., these antibodies Even though PhihccK can be bound at the same time, if the dynamic mutuality Y1 is not shown... C) Viscosity Kawa V-tech, itiotype-anti-idiotype interaction seems to be reasonable. It suggests that. (i.e. j31L) 1 and I31θ2 and ヱ, h　c　G Idiotype between the free fi' (a6) arms that was destroyed because of the vc introduction. 771 that causes itiotype interaction is also good, and 27h L Ip et al d I Q l to n l 03 r! Don't wait for a relationship like this. Sushi However, BIOI and B103 are showing signs of improvement. 1) The notehead of each antibody (for example, 11 PIJ is an epitope empty cycle, It's important to note that Itiotog's Eiji) is regrettable in the 1st month and the 1st month. By becoming a V-box base with a guarded sword, it will be Φ. Complete elucidation of the mechanism In order to do this, there is a need for a summer of experience in history.

Antibody tiger f/I, ii M and serve the small affinity of individual antibodies 2 It conveys two important meanings. g-, the affinity of the monoclonal antibody is the same as above. 1 Birth party〃≦Giving Pillar 0Secondly, in the thread of Hayashi Chi, I will mix the late antibodies and attend. Due to this, there are 4 results for all 9 children with pseudo-assembled young fruits that occur in the polyclonal older brother. Concerning all of the two pairs of monoclonal antibodies in Run 1st munoassay all actual flow, successive mutual f for degree (prevalence) ) All measurements and length. The phase end was measured by two-antibody radioimmunoassay. 1'E (if mixed with 7L, the affinity for the antigen will be 1). Shows G resistance. The plus sign is the antibody on the upper side of the table, which is synergistic with the antibody on the left side of 5. Indicates.

The minus sign indicates a mutually exclusive 1' [and Swallow (゛).

In the dragon, there are 5 strings in the 9th part of the 10 towns.

Well, what do these antibodies do? At the same time, it becomes an antigen at the same time. Because it is not connected in 〃5, the ratio 4J is even higher than I. KoroL et al.'s crude soup When removed, 5 out of 7 assemblies! ll1 katase is required @endnote.

Figure r, n1oi and 4 31 @2 and] 737. O’fill, [i+d101 & antibody B IU For each of 2, 1 heG (-fig) and hLH (=o), which hormones are similar? The logit suppression curves shown for la and are shown in full.

For diagnosis of pregnancy, death, and cancer, this device is equipped with the ability to identify 200 million hormones.

Graph At engineering, BIOI (!: Mixed juice proboscis with f31L12 is more considerate to hLH To serve hcG 170 times more often? Show fo! U7B is BIOI, p; bLf (to. From rough C7, 102 is cut to hCG and has a sensitivity of about 10. after hL) f. Show t.

Therefore, a mixture of antibodies 1+MJ can also be mixed with different antibodies.

The sweetness of the monotalonal antibody increases the affinity for the antigen as described above. 'F-F Note: As I progressed further, I continued my research. Through this research Monoclonal antibodies that have this property are stable between immiscible substances and antigens. Samurai are distinguished from those who do not have the above-mentioned characteristics on the basis of their rudder power, which can form a competitive advantage. Ru. This Chindai 7 is in a state of intense gelatinousness (and has been acting as an extra band). .

L911+glaze contains 1 to 5μ) of hCG and 1 to 5μ of antibody Yjb 8m and the length of 3 + 4 + 5 + 6 and 7% polyacrylamide gels I'm a believer in 111. A constant current of 94 mA is applied to the Ta. 1 addition force method is P, d, 0'Farrell.

(1975) J. Biol. Chem. 250:4007-40211 clj'; The method is essentially the same as the method used to create the original IJ, but with the exception that S O8+Ma1 updated sword λtsuta. Due to the intoxication, the gel was dyed with an appropriate dye.

For the formation of a complex between the monoclonal antibody mixture and the antigen, The emergence of a single, sophisticated monoclonal antibody for the proper antigen assay V This shows that the measurement sensitivity is the same as above.

In addition to the electrokinetic method, the combination of antibodies that produce affinity 1 as above can be determined using gel P increase method. Butyric acid was detected by the extra high molecular weight peak in the chromatography. Niji In detail, an overcharged radioiodinated hcG (0.1 to 10.0 μCi) was applied. 1 to 5 μq of hCG to Pi: mixed with 1 to 10 μ7 of antibody mixture, O,0 15M Herpes buffer, pt47.4-0.9%i”Jact-I A/mIB S In A, 5 ephacryl with a diameter of 1 tx and a width of 24- Chromatography was performed on an S-400 column. B causing affinity ditto The mixture of 101+B102+hCG produced an extra peak, but the mixture of BIOI 10B 1, which did not cause Finitei the same as above. 03+hCG's i-konba proboscis produces an extra beak. Therefore, this method In order, it is important to know which combination of antibodies is effective. I can make a furnace.

Measure the assay to the mark! Use of Tamari monoclonal anti-sweating proboscis that makes sensitivity 7 same as above. Not sure about the purpose? Congratulations, ρS, Fuyuan Ming, to give +j ditto of the annotation. It is also possible to use one bottle of the mixture of The present invention also aims at further application of the mixture for in vivo processing power methods for various conditions. Ruko 8, Contents of correction (1) Pursuant to the provisions of Article 184-5, Paragraph 1 of the Patent Act, which accurately describes the representative of the applicant. Supplement the documents as shown in the attached sheet.

■ Added the translated text on page 29 of the claims, which was typewritten and printed, as shown in the attached document. do. (tt”) @-Two things ψ too・) (4) Power of attorney/corporate personality certificate and translation separately Replenish as per paper.

International search report 794""! 19-500!18Ei (j's) Yo ” Continuation of page 1 0 shots by Neeliff Paul H New York 100, USA 25 New York Haven Avenue 00 @Inventor Moyle William Earl New Jersey, USA 08 854 Pisquitaway River Road 952

Claims (1)

[Claims] 1. Antigen τ of the length of the instep A mixed proboscis of the null antibody, a different antigen site LC, and at least 2 proboscises of different antigens. :Kin's monoclonal antibody Riyatani τ/d young assay 81 formula: i? 9 appropriate Antigens and all monochrome ♂ things in the sweat strips. 2, 24iA Remonoclonal Enemy tzamugibo flood a Mouth kiss 3, cut, Nanamihara,) Bo1 no Pefuchito (: > 9 Ai+: flooded here +31i anti-)iHl list/Created Amino Toshikatsu C that has been added to the peptide Ru 61 Izuku no l Mi no 4 edition i 1-self machine's 7 konsunro. 4. Month 11. Self-horipeftid revenge ceremony, Nobuhito rR Takuo Ko' Natotorohishi (2 Rii = t4 PoL7Jfi-Regi: 'rr'J 1 v (iif; 4Qζ-62 ) 7m 61 course, v5, cut 60 polybegnato Aihara, 0 egg + g #lj is Hol Mo/de ruru 1; djiku no yoke, z6 1 vc5 self-+tv, 'D i B゛ ? /I 1b,j (jl (iself horipenotide 1JT, 7jjL,)i thyroid J lij 7 hormones de ril 1jrJ; F-'s corpse dlvc story I a, aIJ polypeptide 17LIJA2J The sign of the day, the first one is the fart of the people. 8. Square Og self-M efficacy assay weight is 616 self-antigens present in 4 in 1,000 self-sample samples. At least about 10% ρS is enough to form a fixed stand neo for each monochrome A mixture of cavities in 4QI51 of the award of null antibodies. 9. Violet of another antibody [friend of valley antibody against] - iα of about 10':l to about 1:106 1 Claim 1 contains a mixture containing 6 pins; Jl: 10" yoke d9 with 7 konzo proboscises. 11. The shield of the valley antibody against the firefly of another antibody is the ratio of the ridge size of the antibody to the antigen. The mixture J0 as stated in Claim 9 which is the same as the actual purchase 12, 24 stories of monoclonal antibodies and 2. persimmon sweat on the alpha head of gonadotropin, and 1 The antibody introduced in the mixture 013, α-neck & C according to claim 4 which is tri♂ lO2 or AlO3 and the antibody that binds to the finger chain β-chain are B102 or B103. 4U of finger cup binru bill! , 75% of d11v on d12. 14, zz = Group monoclonal antibody t, both of which are human paper hair gonadotes. tj8 in the β-chain of lopin, and the so-called n6 A mixture according to scope 4. ゛Engraving (contents (no changes)) 15. Solid matrix) Antigen containing the mixture according to claim 1 adsorbed on IJ matrix Compositions useful for sensitive assays. 16. Antibiotics containing the mixture according to claim 1 dissolved in a suitably buffered solution. Compositions useful for highly sensitive assays. 17. A method for determining the presence of an antigen in a sample, comprising: The mixture and sample described above are used to form a detectable complex between said mixture and said antigen. A method comprising contacting under suitable conditions to induce formation. 18. A method for determining the presence of human chorionic gonadotropin in a sample, the method comprising: The sample and the mixture according to claim 4, 12.13 or 14, and human chorionic gonadotropin under conditions capable of forming a detectable complex between A method consisting of bringing into contact with