荧光抗体偶联物制备中的技术考量
TECHNICAL CONSIDERATIONS IN THE PREPARATION OF FLUORESCENT-ANTIBODY CONJUGATES.
作者信息
LEWIS V J, JONES W L, BROOKS J B, CHERRY W B
出版信息
Appl Microbiol. 1964 Jul;12(4):343-8. doi: 10.1128/am.12.4.343-348.1964.
Abstract
A comparison was made of (NH(4))(2)SO(4), HCl, ethodin, and ethanol for fractionation of rabbit antiserum prior to conjugation with fluorescein isothiocyanate. Fractionation with the salt was found to be the method of choice from the standpoints of simplicity and recovery of antibody effective in conjugates prepared from the fractions. Effects of pH, temperature, dye-protein ratio, and molarity and type of buffer upon conjugation were studied. These technical factors were adjusted to produce conjugates for Corynebacterium diphtheriae which possessed higher specific titers than did reagents obtained by previously employed techniques.
摘要
在与异硫氰酸荧光素结合之前，对硫酸铵、盐酸、乙锭和乙醇用于兔抗血清分级分离进行了比较。从简便性和从分级分离物制备的结合物中有效抗体的回收率来看，发现用盐进行分级分离是首选方法。研究了pH、温度、染料与蛋白质比例以及缓冲液的摩尔浓度和类型对结合的影响。调整这些技术因素以制备用于白喉棒状杆菌的结合物，这些结合物比以前采用的技术获得的试剂具有更高的特异性滴度。
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