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Study on production of region-specific antibodies against brain-gut hormones

抗脑肠激素区域特异性抗体的制备研究

基本信息

批准号：
63870093
负责人：
YANAIHARA Noboru
金额：
$ 4.16万
依托单位：
University of Shizuoka School of Pharmaceutical Sciences
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1989
项目状态：
已结题

项目摘要

Synthesis of brain-gut hormones: We have synthesized several brain- gut hormones and the related peptides containing 15-20 amino acid residues. Syntheses of these peptides were performed according to solid phase technology using an automated peptide synthesized, Beckman Model 990. A protected peptide chain was constructed on a PAM resin (Applied Biosystems). The purity of the synthetic peptides was proved by amino acid analyses of both acid hydrolysates and AP-M digests of the peptides, analytical C_<18> reverse-phase HPLC and the sequence analysis of the peptide using an automated gas-phase sequencer (Applied Biosystem 470A)., We have synthesized in the present study several new brain-gut hormones such as porcine and rat galanin, helodermin, guinea pig VIP, rat PHI, human glucagon-like peptide-1 (GLP-1), human NPY human and rat proLH-RH-related peptide such as gonadotropin releasing hormone-associated peptide (GAP).Production of antisera against brain-gut hormoned: GLP-1 for productio … More n of GLP-1 specific antisera, synthetic GLP-1(7-22) in porcine thyroglobulin-conjugated form and GLP-1(1-35) in non-conjugated form were selected as immunogens and rabbits were immunized with each of the immunogens. The resulting antisera, anti-GLP-1(7-22) serum RO322 and anti-GLP-1(1-35) serum RO351, were used for the development of radioimmunoassay (RIA)-systems region-specific for GLP-1.Galanin: Since galanin has structural similarities in its C-terminal region to tachykinins, a monoclonal antibody against synthetic porcine galanin(1-15) was successfully produced especially for immunohistochemical study. In fact, this monoclonal antibody was found to recognize specifically galanin-producing cells and did not cross-react with any of tachykinins.Pro-LHRH related peptide: LHRH precursor has been known to contain LHRH and a GAP with 56 amino acid peptide. Using synthetic GAP(34-56) and proLHRH(1-26), we have developed human pro-LHRH carboxy- and amino- terminal specific RIA with the production of anti GAP(34-56) serum(RO242) and anti-proLHRH(1-26) serum (RO620). An antiserum against synthetic rat GAP(28-51) (RO246) was also generated in a rabbits. Less

脑肠激素的合成：我们已经合成了几种脑肠激素和含有15-20个氨基酸残基的相关肽。这些肽的合成根据固相技术使用自动肽合成器Beckman Model 990进行。在PAM树脂（Applied Biosystems）上构建受保护的肽链。通过对肽的酸水解产物和AP-M酶的氨基酸分析、分析C_<18>反相HPLC和使用自动气相测序仪（Applied Biosystem 470 A）的肽序列分析来证明合成肽的纯度。本研究合成了几种新的脑肠激素，如猪和大鼠的甘丙肽（galanin）、helodermin、豚鼠VIP、大鼠PHI、人胰高血糖素样肽-1（GLP-1）、人NPY、人和大鼠的促性腺激素释放肽相关肽（GAP）等，并制备了抗脑肠激素GLP-1的抗血清。 ...更多信息选择n种GLP-1特异性抗血清、猪甲状腺球蛋白结合形式的合成GLP-1（7-22）和非结合形式的GLP-1（1-35）作为免疫原，并用每种免疫原免疫兔子。将所得抗血清，抗GLP-1（7-22）血清RO 322和抗GLP-1（1-35）血清RO 351，用于GLP-1区域特异性放射免疫测定（RIA）系统的开发。甘丙肽：由于甘丙肽在其C-末端区域与速激肽具有结构相似性，因此成功制备了针对合成猪甘丙肽（1-15）的单克隆抗体，特别用于免疫组织化学研究。Pro-LHRH相关肽：已知LHRH前体含有LHRH和56个氨基酸的GAP肽。用人工合成的GAP（34-56）和proLHRH（1-26）建立了人proLHRH羧基和氨基末端特异性RIA，制备了抗GAP（34-56）血清（RO 242）和抗proLHRH（1-26）血清（RO 620）。还在兔中产生了针对合成的大鼠GAP（28-51）（RO 246）的抗血清。少