A rapid quantitative staphylococcal co-agglutination assay. Utilization for the assay of bovine factor VIII-related antigen |
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| Authors: | B V Pandya E P Kirby |
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| Affiliation: | The Wistar Institute of Anatomy and Biology, 36th Street at Spruce, Philadelphia, PA 19104, U.S.A. |
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| Abstract: | A simple and rapid technique to measure bovine factor VIII-related antigen has been developed which utilizes protein A-bearing staphylococci and monospecific rabbit antiserum to bovine factor VIII. Staphylococci coated with a specific antibody agglutinate when they are mixed with the specific antigen. We have used an aggregometer to detect an quantitate the agglutination of the antibody-coated staphylococci. The assay has been optimized with respect to amount of antiserum needed for coating staphylococci, concentration of antibody-coated staphylococci, pH and ionic strength of the assay system, and stirring speed of the aggregometer. The staphylococcal co-agglutination assay as monitored by an aggregometer is at least 10 times more sensitive than the conventional slide agglutination method, and can detect as little as 0.1 microgram/ml of factor VIII antigen. It however, cannot be used to quantitate factor VIII-related antigen in plasma, since plasma contains some components which can non-specifically agglutinate staphylococci. |
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| Keywords: | EDCI 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide SRBC sheep red blood cells RSA rabbit serum albumin KLH keyhole limpet hemocyanin 3-pyridylazo 4-azophthalate IBC antigenic determinant presumably isobutoxycarbonyl group derived from the reaction of protein and isobutylchloroformate TNBS 2,4,6-trinitrobenzene sulfonic acid TNP 2,4,6-trinitrophenyl DNP 2,4-dinitrophenyl PBS phosphate buffered saline BBS 3P-PHPA 3-(3-pyridylazo)-4-hydroxyphenylacetic acid M.D. myristoyl oxidized dextran CFA complete Freund's adjuvant ICFA incomplete Freund's adjuvant HAT hypoxanthine aminopterin thymidine DMEM Dulbecco's modified Eagle's medium PFC plaque forming cells C′ complement FCS fetal calf serum I.P. intraperitoneal all other abbreviations are identified in Table 1 and Fig. 1 |
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