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1
Cold pepsin digestion: a novel method to produce the Fv fragment from human immunoglobulin M.冷胃蛋白酶消化法：一种从人免疫球蛋白M制备Fv片段的新方法。
Proc Natl Acad Sci U S A. 1978 Jun;75(6):2649-53. doi: 10.1073/pnas.75.6.2649.
2
Alternative mechanism of protein A-immunoglobulin interaction the VH-associated reactivity of a monoclonal human IgM.蛋白A与免疫球蛋白相互作用的另一种机制：单克隆人IgM的VH相关反应性
J Immunol. 1985 Aug;135(2):1232-8.
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Production and characterization of an antibody Fv fragment 15N-labeled in the VL domain only.仅在VL结构域进行15N标记的抗体Fv片段的制备与表征。
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4
Noncovalent association of heavy and light chains of human immunoglobulins. IV. The roles of the CH1 and CL domains in idiotypic expression.人免疫球蛋白重链和轻链的非共价结合。IV. CH1和CL结构域在独特型表达中的作用。
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Production of VL and CL fragments from human Bence-Jones kappa chains.从人本斯-琼斯κ链产生VL和CL片段。
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The structural basis for binding of complement by immunoglobulin M.免疫球蛋白M结合补体的结构基础。
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Protein engineering of antibody binding sites: recovery of specific activity in an anti-digoxin single-chain Fv analogue produced in Escherichia coli.抗体结合位点的蛋白质工程：在大肠杆菌中产生的抗地高辛单链Fv类似物中恢复比活性。
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本文引用的文献

1
The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis.通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳测定分子量的可靠性。
J Biol Chem. 1969 Aug 25;244(16):4406-12.
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Macroglobulin structure: variable sequence of light and heavy chains.巨球蛋白结构：轻链和重链的可变序列。
Science. 1970 Jul 3;169(3940):56-9. doi: 10.1126/science.169.3940.56.
3
Immunoglobulin M: pentameric Fcmu fragments released by trypsin at higher temperatures.免疫球蛋白M：在较高温度下由胰蛋白酶释放的五聚体Fcmu片段。
Proc Natl Acad Sci U S A. 1970 Feb;65(2):318-22. doi: 10.1073/pnas.65.2.318.
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Circular dichroism study of the antibody combining site.抗体结合位点的圆二色性研究。
Biochemistry. 1973 Oct 23;12(22):4541-3. doi: 10.1021/bi00746a037.
5
Complete amino acid sequence of the Mu heavy chain of a human IgM immunoglobulin.一种人类IgM免疫球蛋白的μ重链的完整氨基酸序列。
Science. 1973 Oct 19;182(4109):287-91. doi: 10.1126/science.182.4109.287.
6
An unusual papain fragment containing the V H region of an IgG3 myeloma protein.一个含有IgG3骨髓瘤蛋白VH区的异常木瓜蛋白酶片段。
J Immunol. 1972 Sep;109(3):565-9.
7
An active antibody fragment (Fv) composed of the variable portions of heavy and light chains.一种由重链和轻链可变部分组成的活性抗体片段（Fv）。
Biochemistry. 1973 Mar 13;12(6):1130-5. doi: 10.1021/bi00730a018.
8
Localization of antibody-combining sites within the variable portions of heavy and light chains.抗体结合位点在重链和轻链可变区的定位。
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9
Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane.人红细胞膜主要多肽的电泳分析。
Biochemistry. 1971 Jun 22;10(13):2606-17. doi: 10.1021/bi00789a030.
10
The switch point in mu heavy chains of human IgM immunoglobulins.人类IgM免疫球蛋白μ重链中的转换点。
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冷胃蛋白酶消化法：一种从人免疫球蛋白M制备Fv片段的新方法。

Cold pepsin digestion: a novel method to produce the Fv fragment from human immunoglobulin M.

作者信息

Lin L C, Putnam F W

出版信息

Proc Natl Acad Sci U S A. 1978 Jun;75(6):2649-53. doi: 10.1073/pnas.75.6.2649.

DOI:10.1073/pnas.75.6.2649
PMID:351613
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC392620/
Abstract

A strategy for the proteolytic fragmentation of human IgM has been developed. This method is called "cold pepsin digestion" because of its unique feature of achieving restricted peptic cleavages at 4 degrees and pH 4.0. Cold pepsin digestion has been applied successfully to produce an Fv fragment from 14 human IgM proteins. The Fv fragment consists of the heavy chain variable domain (VH) and the light chain variable domain (VL) held together by strong noncovalent interaction. Thus, each Fv fragment contains one intact antigen-binding site and represents the minimal active fragment derivable from an antibody molecule. A series of other structurally and functionally important fragments were also isolated and characterized. Two basic digestion pathways were recognized; these mainly reflect the relative accessibility of five sets of major interdomain cleavage sites.

摘要

已开发出一种用于人免疫球蛋白M（IgM）蛋白水解片段化的策略。由于该方法在4℃和pH 4.0条件下实现有限的胃蛋白酶切割这一独特特性，故称为“冷胃蛋白酶消化”。冷胃蛋白酶消化已成功应用于从14种人IgM蛋白中产生Fv片段。Fv片段由通过强非共价相互作用结合在一起的重链可变结构域（VH）和轻链可变结构域（VL）组成。因此，每个Fv片段包含一个完整的抗原结合位点，代表了可从抗体分子衍生的最小活性片段。还分离并表征了一系列其他结构和功能上重要的片段。识别出了两条基本的消化途径；这些主要反映了五组主要结构域间切割位点的相对可及性。