Hargrave P A, Adamus G, Arendt A, McDowell J H, Wang J, Szaby A, Curtis D, Jackson R W
Exp Eye Res. 1986 Apr;42(4):363-73. doi: 10.1016/0014-4835(86)90030-8.
Antisera and monoclonal antibodies to rhodopsin were examined for their binding specificity to rhodopsin by using peptides from the rhodopsin sequence as competitors for antibody binding to rhodopsin in an enzyme-linked immunoassay. Monoclonal antibodies tested were raised in mice against bovine and rat rhodopsin. Antisera tested were raised in sheep against bovine rhodopsin and in rabbits against human rhodopsin. Peptides were synthesized from the bovine rhodopsin sequences 2-32, 1-12, 13-23, 24-34, 5-11, 231-252 and 331-348 for use as competitors in the immunoassay. A mixture of soluble CNBr peptides, and the purified CNBr peptide representing the sequence 2-39 were also employed. The monoclonal antibodies were all anti-amino-terminal in their binding specificity, although each recognized slightly different regions of the amino terminus. Each of the three antisera was predominantly directed against rhodopsin's amino terminus. We conclude that the amino-terminal 30 or more amino acids, and particularly the amino-terminal 15 amino acids, represent a principal antigenic region of the rhodopsin molecule.
通过在酶联免疫分析中使用来自视紫红质序列的肽作为抗体与视紫红质结合的竞争剂，检测了抗视紫红质抗血清和单克隆抗体对视紫红质的结合特异性。所测试的单克隆抗体是在小鼠体内针对牛和大鼠视紫红质产生的。所测试的抗血清是在绵羊体内针对牛视紫红质产生的，在兔子体内针对人视紫红质产生的。从牛视紫红质序列2 - 32、1 - 12、13 - 23、24 - 34、5 - 11、231 - 252和331 - 348合成肽，用作免疫分析中的竞争剂。还使用了可溶性溴化氰肽混合物以及代表序列2 - 39的纯化溴化氰肽。单克隆抗体在结合特异性上均为抗氨基末端，尽管每个抗体识别氨基末端略有不同的区域。三种抗血清中的每一种主要针对视紫红质的氨基末端。我们得出结论，氨基末端30个或更多氨基酸，特别是氨基末端15个氨基酸，代表视紫红质分子的主要抗原区域。
