Abstract
PREPARATION of antibodies is greatly facilitated by the use of insoluble immuno-adsorbents. We have found that cross-linkage of soluble antigens with formaldehyde readily yields adsorbents having a high antibody capacity and which are insoluble over a wide range of pH. Most experiments have been carried out with an antigen (BSA–CPE) prepared by coupling the diazotized p-amino-phenylether of choline to bovine serum albumin (10 mols/mole). An insoluble polymer of the BSA–CPE was prepared by slowly adding an equal volume of formaldehyde solution (40 per cent; pH 3.1) to a stirred 20 per cent solution of the antigen in 0.15 M sodium chloride. The reaction was allowed to proceed for 1 h at 25° C before the insoluble floccules of antigen polymer were separated by centrifuging. The floccules were washed exhaustively to remove excess formaldehyde with a solution containing 2 M sodium chloride and 0.01 M ammonium chloride adjusted to pH 9.0 with ammonium hydroxide, and finally with 0.15 M sodium chloride.
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Onoue, K., Vagi, Y., and Pressman, D., Immunochemistry, 2, 181 (1965).
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METCALFE, J., MARLOW, H. & BURGEN, A. Immuno-adsorbents of High Capacity. Nature 209, 1142 (1966). https://doi.org/10.1038/2091142a0
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DOI: https://doi.org/10.1038/2091142a0
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