Hey A W, Johnsen J I, Johansen B, Traavik T
Department of Virology, University of Tromsø, Norway.
J Immunol Methods. 1994 Aug 1;173(2):149-56. doi: 10.1016/0022-1759(94)90294-1.
Production of antisera against proteins that are not amenable to easy purification is most efficiently achieved by expressing the protein as a fusion product in bacteria. However, smaller polypeptides may present difficulties, since the majority of the antibodies may be directed against the fusion partner if the whole fusion protein is used as immunogen, while the target peptide alone may be a poor immunogen due to its small size. We have circumvented this problem through the use of two different fusion partners. The first fusion protein is used for priming the immune response and the first boost, while another fusion partner is substituted for the second boost. Five different polypeptides derived from the human polyomavirus BK, ranging in molecular weight from 4400 (39 amino acid residues) to 11,500 (96 amino acid residues), were used to test this approach. The results obtained indicate that this procedure may be very useful in raising antibodies against small antigens.
通过在细菌中表达蛋白质作为融合产物，最有效地实现了针对难以纯化的蛋白质制备抗血清。然而，较小的多肽可能会带来困难，因为如果将整个融合蛋白用作免疫原，大多数抗体可能针对融合伴侣，而单独的目标肽由于其体积小可能是一种较差的免疫原。我们通过使用两种不同的融合伴侣解决了这个问题。第一种融合蛋白用于引发免疫反应和首次加强免疫，而另一种融合伴侣则用于第二次加强免疫。使用了五种源自人多瘤病毒BK的不同多肽，分子量从4400（39个氨基酸残基）到11500（96个氨基酸残基），来测试这种方法。所获得的结果表明，该程序在产生针对小抗原的抗体方面可能非常有用。
