Insights on Monoclonal Antibodies to Kininogens' Heavy Chain Which Influence Kininogens' Binding to Platelets
Insights on Monoclonal Antibodies to Kininogens' Heavy Chain Which Influence Kininogens' Binding to Platelets
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关于影响激肽原与血小板结合的激肽原重链单克隆抗体的见解
DOI：
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发表时间：
1992
影响因子：
6.7
通讯作者：
A. Schmaier
中科院分区：
文献类型：
--
作者：
Yongping Jiang;W. Nawarawong;F. Meloni;A. Schmaier
摘要 低分子量激肽原 (LK) 的纯化结构域可直接用于确定单克隆抗体 (mAb) 的表位，单克隆抗体 (mAb) 已被证明会影响激肽原功能。 LK通过羧甲基-木瓜蛋白酶-Sepharose 4B亲和层析和高岭土吸附从血浆中纯化，并用胰蛋白酶和胰凝乳蛋白酶消化。然后通过凝胶过滤和羧甲基-木瓜蛋白酶-Sepharose 4B 亲和层析分离LK的结构域。使用 LK 重链的纯化结构域，通过酶联免疫吸附测定和免疫印迹确定各种单克隆抗体针对的激肽原重链上的区域。中和激肽原抑制钙蛋白酶的能力的MAb 2B5与结构域2和3交叉反应。发现与激肽原结构域1和2反应的MAb HKH8抑制125I-HK与血小板的结合。在两倍摩尔过量时，mAb HKH8 比更高浓度的 125I-HK 与血小板结合更有效，其中该抗体被证明会导致与血小板的结合增加。或者，即使在高浓度的抗体下，HKH8 F(ab')2 也完全抑制 125I-HK 与血小板的结合。这些研究表明，激肽原重链的纯化结构域可用于快速定位抗体的表位。此外，mAb HKH8 应该是了解激肽原与血小板结合机制的有价值的探针。
Summary Purified domains of low molecular weight kininogen (LK) can be used directly to determine the epitopes of monoclonal antibodies (mAbs) that have been shown to influence kininogen function. LK, purified from plasma by carboxymethyl-papain-Sepharose 4B affinity chromatography and kaolin adsorption, was digested by trypsin and chymotrypsin. The domains of LK were then separated by gel filtration followed by carboxymethyl-papain-Sepharose 4B affinity chromatography. Using the purified domains of LK’s heavy chain, the regions on kininogens' heavy chain which various monoclonal antibodies are directed to were determined by enzyme-linked immunosorbent assay and immunoblotting. MAb 2B5 which neutralized kininogens' ability to inhibit calpain cross-reacted with domains 2 and 3. MAb HKH8 which reacted with kininogens' domain 1 and 2 was found to inhibit 125I-HK binding to platelets. At two-fold molar excess, mAb HKH8 was a better inhibitor of 125I-HK binding to platelets than higher concentrations, where the antibody was shown to cause increased binding to platelets. Alternatively, HKH8 F(ab')2 completely inhibited 125I-HK binding to platelets even at high concentrations of antibody. These studies indicate that purified domains of kininogens' heavy chain can be used to rapidly localize epitopes for antibodies. Further, mAb HKH8 should be a valuable probe to understand the mechanisms of kininogens' binding to platelets.
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α2-硫醇蛋白酶抑制剂的人 cDNA 的分离及其与低分子量激肽原的同一性。
影响因子：
2.9
作者：
Ohkubo,I;Kurachi,K;Takasawa,T;Shiokawa,H;Sasaki,M
通讯作者：
Sasaki,M
通过针对高分子量激肽原每条链的单克隆抗体研究来确定其双功能特性。
DOI：
--
发表时间：
1987
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Schmaier,AH;Schutsky,D;Farber,A;Silver,LD;Bradford,HN;Colman,RW
通讯作者：
Colman,RW
高分子量激肽原是血小板钙蛋白酶的抑制剂。
DOI：
10.1172/jci112472
发表时间：
1986
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
Schmaier,AH;Bradford,H;Silver,LD;Farber,A;Scott,CF;Schutsky,D;Colman,RW
通讯作者：
Colman,RW
影响因子：
7.5
作者：
A. Schmaier;L. Silver;A. Adams;G. Fischer;P. Munoz;L. Vroman;R. Colman
通讯作者：
A. Schmaier;L. Silver;A. Adams;G. Fischer;P. Munoz;L. Vroman;R. Colman
人高分子量激肽原的裂解显着增强其凝血活性。
DOI：
10.1172/jci111319
发表时间：
1984
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
Scott,CF;Silver,LD;Schapira,M;Colman,RW
通讯作者：
Colman,RW