COMPARISON OF 2 PRECIPITATING DERIVATIVES OF RABBIT ANTIBODY - FRAGMENT I DIMER AND PRODUCT OF PEPSIN DIGESTION
COMPARISON OF 2 PRECIPITATING DERIVATIVES OF RABBIT ANTIBODY - FRAGMENT I DIMER AND PRODUCT OF PEPSIN DIGESTION
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DOI：
10.1021/bi00881a024
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发表时间：
1965-01-01
期刊：
影响因子：
2.9
通讯作者：
CEBRA, JJ
中科院分区：
文献类型：
--
作者：
JAQUET, H;CEBRA, JJ
本报告涉及“片段I二聚体”，产品的连续行动的不溶性木瓜蛋白酶和十二烷基硫酸盐对兔γ-球蛋白，和“胃蛋白酶产品”，产品的详尽胃蛋白酶蛋白水解的γ-球蛋白。报道了制备这两种免疫学二价产物的改进方法，这些产物不含任何可检测的与片段HL相关的抗原决定簇污染。片段I二聚体的测量S20，W和D20，w分别为4.96 S和5.63 × 10-7 cm 2/sec，胃蛋白酶产物的测量S20，W和D20，w分别为4.86 S和5.11 × 10 -7 cm 2/sec。片段I二聚体的mw sD为84，000，胃蛋白酶产物的mw sD为90，700。片段I二聚体和胃蛋白酶产物通过许多其他标准进行比较，包括抗体活性、抗原决定簇的含量、琼脂凝胶中的电泳迁移率以及通过单价片段的再结合还原后重新形成的能力。这两种产品的差异最大时，比较后一个标准。通过胃蛋白酶产物的还原产生的片段以良好的产率重组，而在相同条件下片段I二聚体的两个部分不重组。在胃蛋白酶蛋白水解之前，或在胃蛋白酶产物还原之前或之后，用insolbuel木瓜蛋白酶进行短暂处理，导致单价片段在标准条件下重组的能力丧失。
This report concerns "fragment I dimer", the product of the consecutive action of insoluble papain and dodecylsulfate on rabbit gamma-globulin, and "pepsin product," the product of exhaustive pepsin proteolysis of gamma-globulin. Improved procedures are reported for the preparation of both these immunologically divalent products free of any detectable contamination with antigenic determinants associated with fragment HL The measured S20, W, and D20, w for fragment I dimer were 4.96 S and 5.63 x 10-7 cm2/sec, and for pepsin product were 4.86 S and 5.11 x lO-7 cm2/sec. The mw sD of fragment I dimer was 84,000 and of pepsin product was 90,700. The fragment I dimer and pepsin product were compared by a number of other criteria, including antibody activity, content of antigenic determinants, electro-phoretic mobility in agar gel, and ability to re-form after reduction by reassociation of univalent fragments. The two products differed most when compared by the latter criterion. The fragments produced by reduction of pepsin product recombined in good yield while under the same conditions the two moieties of fragment I dimer did not. Brief treatment with insolbuel papain, either before pepsin proteolysis, or before or after reduction of the pepsin product, resulted in loss of ability of the univalent fragments to recombine under standard conditions.