Yukawa N, Matsuda H, Seo Y, Suetomi K, Takahama K
Department of Legal Medicine, Miyazaki Medical College, Japan.
Bioconjug Chem. 1994 May-Jun;5(3):273-7. doi: 10.1021/bc00027a014.
A method for peroxidase labeling of the monomeric subunit (IgMs) of ABO blood group specific mouse monoclonal IgM is described. IgM was purified from a commercial monoclonal anti-B blood grouping reagent by a combination of salt precipitation, euglobulin precipitation, and gel filtration. IgM was mildly reduced with L-cysteine to yield SH-bearing IgMs. Finally, IgMs was conjugated to horseradish peroxidase, into which SH-reacting maleimide groups had been introduced using N-succinimidyl 6-maleimidohexanoate, through the selective reaction between SH of IgMs and maleimide groups of peroxidase.
描述了一种对ABO血型特异性小鼠单克隆IgM的单体亚基（IgMs）进行过氧化物酶标记的方法。通过盐沉淀、优球蛋白沉淀和凝胶过滤相结合的方式，从市售的抗B血型单克隆试剂中纯化IgM。用L-半胱氨酸对IgM进行温和还原，以产生带有SH基的IgMs。最后，通过IgMs的SH基与过氧化物酶的马来酰亚胺基团之间的选择性反应，将IgMs与已使用N-琥珀酰亚胺基6-马来酰亚胺基己酸引入SH反应性马来酰亚胺基团的辣根过氧化物酶偶联。
