Fasciglione G F, Marini S, Bannister J V, Giardina B
Department of Experimental Medicine and Biochemical Sciences, University of Rome Tor Vergata, Italy.
Hybridoma. 1996 Feb;15(1):1-9. doi: 10.1089/hyb.1996.15.1.
The coupling of haptens to carrier proteins is required for the generation of antihapten antibody response. To date, different polypeptides (such as gelatin, albumin, and keyhole limpet hemocyanin) have been used to increase the immune response against haptens. Different methods of conjugation have also been used to increase the specificity of the antibody response. Our results, obtained through an analysis of the antibody response at both the polyclonal and the monoclonal levels, show that carrier proteins specifically modulate the antibody titer, the specificity of the response, the fusion efficiency, and the number of specific clones. Moreover, the affinity constants of both serum and randomly selected hybridomas directed against different haptens have been found to be exclusively modulated by carrier proteins. In addition, the results show that the carrier protein should be selected considering its possible physicochemical interaction with haptens. Data obtained suggested that any carrier is suitable for hydrophilic haptens, while the choice of the carrier for hydrophobic haptens is critical in obtaining a specific immune response. The use of specific carriers allowed the production of highly specific antibodies: even IgMs, obtained by using these carriers, were able to specifically react with hydrophobic haptens; the case of some IgM monoclonal antibodies specifically reacting with protoporphyrin IX is reported.
半抗原与载体蛋白偶联是产生抗半抗原抗体反应所必需的。迄今为止，不同的多肽（如明胶、白蛋白和钥孔血蓝蛋白）已被用于增强对半抗原的免疫反应。不同的偶联方法也被用于提高抗体反应的特异性。我们通过对多克隆和单克隆水平的抗体反应进行分析得出的结果表明，载体蛋白可特异性调节抗体效价、反应特异性、融合效率以及特异性克隆的数量。此外，已发现针对不同半抗原的血清和随机选择的杂交瘤的亲和常数完全由载体蛋白调节。此外，结果表明应根据载体蛋白与半抗原可能的物理化学相互作用来选择载体蛋白。所获得的数据表明，任何载体都适用于亲水性半抗原，而对于疏水性半抗原，载体的选择对于获得特异性免疫反应至关重要。使用特定载体能够产生高度特异性的抗体：即使是通过使用这些载体获得的IgM，也能够与疏水性半抗原特异性反应；报道了一些IgM单克隆抗体与原卟啉IX特异性反应的情况。
