Place J D, Schroeder H R
J Immunol Methods. 1982;48(2):251-60. doi: 10.1016/0022-1759(82)90199-5.
Serum samples were found to be capable of desorbing as much as 40% of the antibody to hepatitis B surface antigen (anti-HBsAg) adsorbed to plastic surfaces. This previously unreported loss could affect the accuracy of the assay, so chemical fixation was examined as a means for preventing antibody desorption during a 'sandwich' radioimmunoassay for HBsAg. Methods for fixing the anti-HBsAg were developed with glutaraldehyde and ethylchloroformate. Both methods prevented antibody desorption from polyvinylchloride and polystyrene without affecting immunoreactivity in radioimmunoassay. A combined glutaraldehyde-ethylchloroformate method resulted in stronger fixation that fully resisted the sera that caused the greatest desorption. It was found that only polymerized glutaraldehyde fixed anti-HBsAg to plastic; the monomer was ineffective. Anti-HBsAg fixed microtiter plates could be stored for at least 4 weeks without loss of sensitivity in radioimmunoassays. These methods could be adapted for use in other assays where the prevention of protein desorption from the solid phase is an important consideration.
研究发现，血清样本能够解吸出吸附在塑料表面的高达40%的乙型肝炎表面抗原抗体（抗-HBsAg）。这种此前未报道的损失可能会影响检测的准确性，因此对化学固定作为一种在HBsAg的“夹心”放射免疫分析中防止抗体解吸的方法进行了研究。用戊二醛和氯甲酸乙酯开发了固定抗-HBsAg的方法。这两种方法都能防止抗体从聚氯乙烯和聚苯乙烯上解吸，且不影响放射免疫分析中的免疫反应性。戊二醛-氯甲酸乙酯联合方法产生更强的固定效果，能完全抵抗导致最大解吸的血清。研究发现，只有聚合的戊二醛能将抗-HBsAg固定到塑料上；单体无效。固定有抗-HBsAg的微量滴定板在放射免疫分析中可储存至少4周而不丧失敏感性。这些方法可适用于其他检测，其中防止蛋白质从固相解吸是一个重要的考虑因素。
