Peterman J H, Tarcha P J, Chu V P, Butler J E
Department of Microbiology, University of Iowa, Iowa City 52242.
J Immunol Methods. 1988 Jul 22;111(2):271-5. doi: 10.1016/0022-1759(88)90136-6.
The antigen capture capacity of antibodies covalently immobilized on injection-molded polystyrene beads was evaluated. Bromoacetyl groups on the bead surfaces rendered them reactive to protein nucleophilic groups. The bromoacetyl surface exhibited up to a ten-fold greater capacity for protein compared to unmodified polystyrene, with no detectable dissociation such as occurs with simple adsorption. Biotinylated anti-fluorescein was immobilized on this surface both through direct covalent attachment and indirectly via streptavidin, which was first covalently attached to the bead. Comparisons of the resulting biological activity, normalized to the amount of anti-fluorescein on the bead, were made between the attachment methods and simple passive adsorption. The presence of the streptavidin spacer on the bromoacetyl surfaces improved the antigen capture capacity of antifluorescein, for fluoresyl-albumin by 45% compared to direct covalent linkage of the antibody to modified polystyrene and by 160% relative to antibody adsorbed on unmodified polystyrene.
评估了共价固定在注塑聚苯乙烯珠上的抗体的抗原捕获能力。珠表面的溴乙酰基使其对蛋白质亲核基团具有反应性。与未改性的聚苯乙烯相比，溴乙酰表面对蛋白质的结合能力高达10倍，且未检测到如简单吸附那样的解离现象。生物素化的抗荧光素通过直接共价连接以及先将链霉亲和素共价连接到珠子上再间接固定在该表面上。对两种固定方法以及简单被动吸附所得的生物活性（以珠子上抗荧光素的量进行归一化）进行了比较。溴乙酰表面上链霉亲和素间隔物的存在提高了抗荧光素对荧光酰化白蛋白的抗原捕获能力，与抗体直接共价连接到改性聚苯乙烯相比提高了45%，相对于吸附在未改性聚苯乙烯上的抗体提高了160%。
