CS231763B1 - Preparation method of agent for binding of antisubstances - Google Patents
Preparation method of agent for binding of antisubstances Download PDFInfo
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- CS231763B1 CS231763B1 CS82194A CS19482A CS231763B1 CS 231763 B1 CS231763 B1 CS 231763B1 CS 82194 A CS82194 A CS 82194A CS 19482 A CS19482 A CS 19482A CS 231763 B1 CS231763 B1 CS 231763B1
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- deoxyribonucleoprotein
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- 238000002360 preparation method Methods 0.000 title description 11
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 24
- 239000011780 sodium chloride Substances 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 108091023046 Deoxyribonucleoprotein Proteins 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 210000001541 thymus gland Anatomy 0.000 claims description 4
- 238000010257 thawing Methods 0.000 claims description 3
- 208000035404 Autolysis Diseases 0.000 claims description 2
- 206010057248 Cell death Diseases 0.000 claims description 2
- 230000000694 effects Effects 0.000 claims description 2
- 239000000463 material Substances 0.000 claims description 2
- 230000028043 self proteolysis Effects 0.000 claims description 2
- 230000000391 smoking effect Effects 0.000 claims description 2
- 230000029142 excretion Effects 0.000 claims 1
- 239000002504 physiological saline solution Substances 0.000 claims 1
- 239000000243 solution Substances 0.000 claims 1
- 238000012360 testing method Methods 0.000 description 13
- 108090000790 Enzymes Proteins 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 5
- 239000007790 solid phase Substances 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 239000011230 binding agent Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 201000000596 systemic lupus erythematosus Diseases 0.000 description 3
- 108010077544 Chromatin Proteins 0.000 description 2
- 230000004520 agglutination Effects 0.000 description 2
- 238000003149 assay kit Methods 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 210000003483 chromatin Anatomy 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 238000003018 immunoassay Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000009257 reactivity Effects 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000013049 sediment Substances 0.000 description 2
- 238000000527 sonication Methods 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 230000008961 swelling Effects 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 102000053602 DNA Human genes 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 238000006424 Flood reaction Methods 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 102000001554 Hemoglobins Human genes 0.000 description 1
- 108010054147 Hemoglobins Proteins 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 108010059724 Micrococcal Nuclease Proteins 0.000 description 1
- 108010061100 Nucleoproteins Proteins 0.000 description 1
- 102000011931 Nucleoproteins Human genes 0.000 description 1
- 108010047956 Nucleosomes Proteins 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 240000002657 Thymus vulgaris Species 0.000 description 1
- 235000007303 Thymus vulgaris Nutrition 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 206010061592 cardiac fibrillation Diseases 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 235000019365 chlortetracycline Nutrition 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000012636 effector Substances 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 230000002600 fibrillogenic effect Effects 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 238000000265 homogenisation Methods 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 239000004816 latex Substances 0.000 description 1
- 229920000126 latex Polymers 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 206010025135 lupus erythematosus Diseases 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 210000001623 nucleosome Anatomy 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000013014 purified material Substances 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 1
- 229940038773 trisodium citrate Drugs 0.000 description 1
- 239000013585 weight reducing agent Substances 0.000 description 1
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- Peptides Or Proteins (AREA)
Description
Vynález sa týká sposobu přípravy prostriedku pre viazanie protilátok, ktorý je určený najma pre diagnózu systémového lupus erythematosus.The invention relates to a method for the preparation of an antibody binding agent, in particular for the diagnosis of systemic lupus erythematosus.
Doteraijšie testy pre diagnózu systémového lupus erythematosus vychádzajú z objavu tzv. LE-bunkového fenoménu, resp. testu Hargraves Μ. M., Richmond H., Morton R., Proč. Mayo Glin. 23, 25 až 28, 1948; o tomto fenoméne sa dokázalo, že v ňom ako éfektor účinkujú protilátky proti deoxyriboinukleoproteínu, povodně· označené ako LE-faktory. Je známy rad testov, ktoré tento fakt zúžitkovali a najviac sa rozšířili aglutinačné testy. Vačšinou však tieto testy nespíňajú nároky praxe, najma pokia! ide o citlivost. Aglutinačné testy preferuji! len určitú část protilátok a bývajú pozitivně v 14 až 86 % sér, podlá koncentrácie protilátok, ktoré tieto obsahuji! Chapman J. C., Amer. j. med. Technol. 42, 154 až 157, 1978. V testoch, ktoré sú obchodné dostupné sa používajú latexové častíce, potiahnuté deoxyribonukleoproteínom: firemná literatura Hyland, Lederle, Fisher.Current tests for the diagnosis of systemic lupus erythematosus are based on the discovery of so-called. LE-cell phenomenon, resp. test Hargraves Μ. M., Richmond H., Morton R., Proc. Mayo Glin. 23, 25-28 (1948); This phenomenon has been shown to act as an effector in anti-deoxyriboinucleoprotein floods, referred to as LE-factors. There are a number of tests that have taken advantage of this fact and the most widespread are agglutination tests. In general, however, these tests do not meet the demands of practice, especially if! it is sensitivity. I prefer agglutination tests! only a certain portion of the antibodies and are positively present in 14 to 86% of the sera, depending on the concentration of the antibodies containing these! Chapman J.C., Amer. j. med. Technol. 42, 154-157, 1978. In commercially available tests, latex particles coated with deoxyribonucleoprotein are used: the company literature Hyland, Lederle, Fisher.
Deoxyribonuklooproteín je zložka chromatínu a na jeho přípravu sa využívá jeho nerozpustnost pri fyziologickej iónovej sile, t. j. pri koncentrácii NaCl 0,15 mol. . dm-3 a relativná rozpustnost alebo dispergovatelnosť pri nízkej alebo vyššej iónovej sile. Nedosjtatok tohoto klasického postupu je tendencia deoxyribonukleoproteínu vytvárat iréverzibilne nerozpustný gél, a to už pri relativné nízkej koncentrácii, nad 0,8 g . dm-3 deoxyribonukleoproteínu, ďalej pri prezrážaní alkoholom alebo solarni, a tiež keď sa predčistený materiál nechá dlho stát a homogenizácia nie je dost intenzívna. To sa v súčasnosti ohchádza na základe poznaitku, že chromatín je zložený z menších podjednotiek — nukleozómov — a tieto je možné získat na rozdiel od predchádzajúcich mechanických postupov, působením enzýmov, ako napr. mikrokokovej nukleázy alebo extrakciou při selektívnej koncentrácii niektorých solí Noll M., Thomas J. O., Kornberg R., Science 187,Deoxyribonuclooprotein is a component of chromatin and is used for its preparation by its insolubility at physiological ionic strength, ie at the concentration of NaCl 0.15 mol. . dm -3 and the relative solubility or dispersibility at low or higher ionic strength. A shortcoming of this classical procedure is the tendency of deoxyribonucleoprotein to form an irreversibly insoluble gel, even at a relatively low concentration, above 0.8 g. dm -3 deoxyribonucleoprotein, followed by alcohol or solar precipitation, and also when the pre-purified material is allowed to stand for a long time and homogenization is not intense enough. This is currently being appreciated by the notion that chromatin is composed of smaller subunits - nucleosomes - and these can be obtained, in contrast to previous mechanical procedures, by enzymes such as e.g. micrococcal nuclease or extraction at a selective concentration of some salts of Noll M., Thomas JO, Kornberg R., Science 187,
1203, 1975; Fenske a spol., Acta med. biol. germ. 39, 343 až 354, 1980. Tieto postupy sú zložité, nákladné a výtažky bývajú malé, menšie ako 25 % východzieho materiálu. Takéto přípravky slúžia hlavně pre badatelské ciele.1203, 1975; Fenske et al., Acta med. Biol. germ. 39, 343-354 (1980). These processes are complex, expensive, and yields are small, less than 25% of the starting material. Such preparations are mainly used for research purposes.
Uvedené nevýhody odstraňuje spósob přípravy prostriedku pre viazanie protilátok, tvořeného deoxyribonukleoproteínom, pri ktorého príprave sa vychádza z teíacieho týmusu, jadier kuřácích erytrocytov alebo z materiálu s hmotnostným zlomkom jadier nad 0,5, vyčištěného opakovanou extrakciou fyziologickým roztokom. Podstata vynálezu spočívá v zbotnaní zvyšku a autolýze vo vodě alebo roztoku chloridu sodného v koncontrácii do 0,01 mol. dm-3 počas 8—48 hodin, pri teplote 0 až 10 °C a v 2 až 5-násobnoim podrobení vibračnému účinku s frekvenciou 10 až 30 kHz pri výkone 50 až 1300 W pri teplote 0 až 10 °C. Získaný materiál sa vysuší mrazovou subllmáciou a získá sa stabilný preparát vo formě bielej, páperovitej látky, dobré rozpustnej v roztokoch solí i vo· vodě. Použitelnost takto připraveného prostriedku na viazanie protilátok, LE-faktorov, jo možná na základe poznatku, že aj napriek enzýmovej a vlbračnej degradácii a zníženiu molekulovej hmotnosti si zachovává potřebné anti'génne determinanty.These disadvantages are overcome by a method of preparing an antibody binding agent consisting of a deoxyribonucleoprotein starting from a calf thymus, erythrocyte-smoking cores or a core mass fraction above 0.5, purified by repeated extraction with saline. SUMMARY OF THE INVENTION The invention is based on swelling of the residue and autolysis in water or sodium chloride solution in a concentration up to 0.01 mol. dm -3 for 8-48 hours, at a temperature of 0 to 10 ° C and a 2 to 5-fold vibration effect at a frequency of 10 to 30 kHz at an output of 50 to 1300 W at a temperature of 0 to 10 ° C. The material obtained is freeze-dried and a stable preparation is obtained in the form of a white, fluffy substance which is soluble in both salt and water solutions. The usefulness of the antibody-LE-binding agent composition thus prepared may be based on the knowledge that despite the enzyme and fibrillation degradation and molecular weight reduction, it retains the necessary anti-gene determinants.
Výhodou tohoto sposobu přípravy je hlavně to, že sa dá uskutočniť jednoducho a nenákladné. V laboratómych podmienkach sa dá připravit také množstvo substancie, aké je potřebné na jej priemyselné využitie.The main advantage of this method of preparation is that it can be carried out simply and inexpensively. Under laboratory conditions, the amount of substance required for industrial use can be prepared.
K dókazu účinku slúži nasledujúca tabulka, udáyajúca výsledky testovania sér chorých s pozitívnym LE-bumkovým testom, 'ktorý sa v tomto případe móže považovat za refforenčný. Analýzy sa vykonali testovacími súpravami Hyland, Lederle a pomocou prostriedku podlá vynálezu, enzýmovou imunoanafýzou na detekciu protilátok proti deoxyribonukleoproteínu triedy IgG (IgG— —am'ti—DNP).The following table shows the results of testing the sera of patients with a positive LE-cell test, which in this case can be considered to be reflexive. The assays were performed with Hyland, Lederle assay kits and by means of the composition of the invention, by enzyme immunoassay for the detection of IgG class anti-deoxyribonucleoprotein (IgG-am'ti-DNP) antibodies.
Tabulkatable
Tieto výsledky dokazujú, že prostriedok podlá vynálezu je najúčinnejší v porovnaní s reíerenčným testom, v zmysle zhody v pozitivitě oboch testov.These results demonstrate that the composition of the invention is most effective in comparison with the referral test, in terms of consistency in the positivity of both assays.
Příprava deoxyrlbonuklooproteínu známými postupmi vedře k malým výťažkom produktu, ktorý je limitované rozpustný. Běžný postup spočívá v homogenizácii tkaniva, alebo izolovaných jadier v štandardnom solinom roztoku 0,15 mol. dm'3 NaCl, čo sa opakuje dotial', kým sa už neextrahuje signifikantně množstvo proteinu. Nerozpustný zvyšok isa potom rýchlo homogenizuje v tisíc-násobnom přebytku slabého solného roztoku, 0,015 mol. dim-3 NaCl a menej a centriíuguje pri vysokom zrychlení 15 000 . g a viac. Typicky sa z 10 g íelacieho týmusu vyťaží 0,20' až 0,50 g deoxyribonukleoproteínu.The preparation of the deoxyrone carbonucoprotein by known methods leads to low yields of the product which is of limited solubility. A common procedure is to homogenize tissue or isolated cores in a standard solin solution of 0.15 mol. dm ' 3 NaCl, which is repeated until a significant amount of protein is no longer extracted. The insoluble residue is then rapidly homogenized in a 1000-fold excess of weak saline, 0.015 mol. dim -3 NaCl and less and centrifuges at a high acceleration of 15,000. g and more. Typically, 0.20-0.50 g of deoxyribonucleoprotein are recovered from 10 g of the thymus thymus.
Postup podlá vynálezu je jednoduchý a přitom účinnější. Poskytuje vačšie množstvo isubstancie, s výhodnějšími vlastnosťami. Postup přípravy vynálezu jo ďalej objasněný ina príkladoch, ktoré ale jeho rozsah ani neobmedzujú ani nevyčerpávajú.The process according to the invention is simple yet more efficient. It provides more isubstance, with more advantageous properties. The process of preparation of the invention is further elucidated by other examples, which are not to be construed as limiting or exhaustive.
Přikladl g tefacieho týmusu, ktorý bol predbežne dokladné extrahovaný štandardným solným roztokom, obsahujúcim 0,15 mol. dm-3 NaCl, sa dispergoval pomocou kuchyňského mixéru v 250 ml 0,01 mol. dm-3 NaCl a gél sa nechal stát v chlade pri 5 °C 10 až 18 hod, kým nemabotnal. Gél sa potom dispergoval pomocou injekčnej striekačky s trubičkou 0 2,0 až 2,5 mm a podrobil sonikovaniu pri výkone 100 W a frekvencii 20 kHz. Teplota nesmie vystúpiť na 10 ’C. Po scentrifugovaní pri 15 000 g počas 1 hod, a 5 °C sa sediment podrobil rovnakému sonikovaniu opakované, až sa v sedimente nenachádzala deoxyribonukleová kyselina. Extrakty sa zlyofilizovali a vyťazilo sa tak spolu 1,7 g preparátu. Tento sa dobré rozpúšťa vo vodě alebo v solných roztokoch. Roztoky majú charakteristické spektrum v ultrafialovej oblasti, kde vykazujú Amax = — 257 nm a Amin = 237 inm. Poměr absorbancií pri 260 a 280 nm je 1,3 až 1,5 v štyroch frakiciách a hmotnostný poměr proteín/DNA bol 1,0 až 1,6.Example 1 g of the thawing thymus, which was pre-extracted with a standard saline solution containing 0.15 mol. dm -3 NaCl, was dispersed with a kitchen mixer in 250 ml of 0.01 mol. dm -3 NaCl and the gel was allowed to stand at 5 ° C for 10-18 hours until it swelled. The gel was then dispersed using a syringe with a 2.0-2.5 mm tube and sonicated at 100 W at a frequency of 20 kHz. The temperature must not rise to 10 ° C. After centrifugation at 15,000 g for 1 hour, and 5 ° C, the sediment was subjected to the same sonication repeatedly until no deoxyribonucleic acid was present in the sediment. The extracts were lyophilized to give a total of 1.7 g of the preparation. This dissolves well in water or in saline solutions. The solutions have a characteristic spectrum in the ultraviolet region where they exhibit λ max = - 257 nm and λ min = 237 inm. The absorbance ratio at 260 and 280 nm is 1.3 to 1.5 in four fractions and the protein / DNA weight ratio was 1.0 to 1.6.
Na přípravu je výhodné použit tkanivá bohaté na obsah deoxyribonukleoproteínu a vedla týmusov sú osobitne výhodné aj jadrá z kuřácích erytrocytov, ktoré sa dajú 1'ahko připravit v čistom stave.Deoxyribonucleoprotein-rich tissues are preferred and, in addition to thymes, kernels of smoking erythrocytes, which are easy to prepare in a pure state, are particularly preferred.
Příklad 2 , Asi 150 ml kuracej krvi sa premylo 4 rázy fyziologickým roztokom a podrobilo lýže v 0,01 rool.dim-3 CaCl2. Jadrá sa zachytili scentrifugovaním lyzátu pri nízkých obrátkách a premyli sa štandardným solným roztokom až do zjavného odstránenia hemoglobinu. Jadrá sa narušili dvojnásobným zmrazením a rozmražením a podrobili sa zbotnaniu v 100 ml vody počas 18 hod. Nato sa dispergovali a podrobili sonikovaniu pri 100 W a 20 kHz pri tepláte do 10 °C vždy 1 min, načo sa stočili pri 15 000 g 1 hod. a 5 stupňov Celsia a zvyšok sa opat podrobil sonikovaniu, Po 5 sonikáciách sa prakticky všeíok nukleoproteín jadíer previedol do roztoku a po mrazovej sublimácii sa získalo 1,35 g 'bielej páperoviíej látky. Rozpúšťal sa dobré vo vodě i v solných roztokoch s charakteristickým spekírom v ultrafialové] oblasti. Poměr absorbancií pri 260 a 280 nm bol 1,05 až 1,4 a hmotnostný poměr protein,/DNA 1,0 až 1,7.Example 2: About 150 ml of chicken blood was washed 4 times with saline and lysed in 0.01 rool.dim -3 CaCl 2 . The nuclei were collected by centrifugation of the lysate at low speed and washed with standard saline until the hemoglobin was evidently removed. The cores were disrupted by double freezing and thawing and subjected to swelling in 100 ml of water for 18 hours. They were then dispersed and sonicated at 100 W and 20 kHz at a temperature of up to 10 ° C for 1 min each, after which they were centrifuged at 15,000 g for 1 hour. and 5 degrees Celsius and the remainder was sonicated. After 5 sonications, virtually all of the nucleoprotein of the livers were dissolved and freeze-dried to give 1.35 g ' It dissolves well in water and in saline solutions with a characteristic spectrum in the ultraviolet range. The absorbance ratio at 260 and 280 nm was 1.05 to 1.4 and the protein / DNA weight ratio was 1.0 to 1.7.
V priebehu niekolkých dní sa dá připravit týmto postupom také množstvo substancie, ktoré vystačí pre priemyslové zhotovenie testovacích súprav na screening systémového lupus erytíhematosus. Prosíriedok je dlhodobe skladovatelný v chladničke pri teplote pod 10 °C. Preparáty skladované 2 roky sa dali použit v teste. Před přípravou testovacích skúmaviek alebo mikrodosiek sa prostriedok rozpustí vo vodě a jeho koncentrácl-a sa nastaví výhodné tak, že v sol'nom roztoku 0,15 mol. dm“3 NaCl a 0,015 mol. dm-3 trinátriumcitrát dává absorbanciu pri 260 nm 0,5. Skúmavky, mikrodosky, všeobecne peviná fáza, sú výhodné z polystyrénu a naplnia sa roztokom prostriedku tak, aby sa získala dostatočná aktívna plocha.Within a few days, the amount of substance that is sufficient for the industrial production of test kits for screening systemic lupus erythhematosus can be prepared by this procedure. The product is stored in a refrigerator below 10 ° C for a long time. Preparations stored for 2 years could be used in the assay. Prior to the preparation of test tubes or microplates, the composition is dissolved in water and its concentrates are preferably set so that in a saline solution of 0.15 mol. dm 3 NaCl and 0.015 mol. dm -3 trisodium citrate gives an absorbance at 260 nm of 0.5. Tubes, microplates, generally the solid phase, are preferably polystyrene and filled with a solution of the composition so as to obtain a sufficient active area.
Pláštenle, aktivovanie pevnej fázy prebehne za 15 až 18 hod. pri 5 °C, načo sa roztok odsaje a pevná fáza sa opakované prernyje vodou. Testovanie sér sa uskuteční naplněním vhodné riedeného séra, ktoré sa po inkubácii odstráni a pevná fáza sa opat dokladné přeplácíme. Viazané protilátky sa hapokon odhalia inkubáciou, pomocou amtiglobulínového konjugátu, označeného výhodné enzýmom, ktorého farebný produkt je úměrný koncentrácii viazaného konjugátu a tým aj protilátky. Rozsah reaktivity testovaného séra sa može výhodné porovnat s reaktivitami pozítívneho a negativného referenčného séra. Využitie prostriedku v enzýmoimunologickom teste umožňuje špecifický a citlivý screening chorých so systémovým lupus erythematosus, s možnoisťou kvantitativného určenia protilátek v roznych imunoglobulínových triedach.Tires, solid phase activation takes place in 15-18 hours. at 5 ° C, the solution is filtered off with suction and the solid phase is repeatedly rinsed with water. Serum testing is performed by filling with a suitable diluted serum, which is removed after incubation and the solid phase is carefully repaid. Bound antibodies are hapokone detected by incubation with an amtiglobulin conjugate, labeled with a preferred enzyme, whose color product is proportional to the concentration of bound conjugate and hence of the antibody. The extent of reactivity of the test serum can advantageously be compared with the reactivities of the positive and negative reference sera. The use of the composition in the enzyme immunoassay enables specific and sensitive screening of patients with systemic lupus erythematosus, with the possibility of quantitatively determining antibodies in various immunoglobulin classes.
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| Application Number | Priority Date | Filing Date | Title |
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| CS82194A CS231763B1 (en) | 1982-01-11 | 1982-01-11 | Preparation method of agent for binding of antisubstances |
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| CS82194A CS231763B1 (en) | 1982-01-11 | 1982-01-11 | Preparation method of agent for binding of antisubstances |
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| CS231763B1 true CS231763B1 (en) | 1984-12-14 |
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