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

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。这些肽的合成是根据固相技术进行的，使用自动合成的肽，贝克曼模型990。在PAM树脂（Applied Biosystems）上构建了保护肽链。合成肽的纯度通过酸水解物和AP-M酶切物的氨基酸分析、C_<18>反相高效液相色谱分析和自动气相测序仪（Applied Biosystem 470A）的序列分析得到证实。本研究合成了猪、大鼠甘丙肽、helodermin、豚鼠VIP、大鼠PHI、人胰高血糖素样肽-1 （GLP-1）、人NPY、人、大鼠prolh - rh相关肽、促性腺激素释放激素相关肽（GAP）等几种新型脑肠激素。选择GLP-1特异性抗血清、合成GLP-1（7-22）型猪甲状腺球蛋白结合型和合成GLP-1（1-35）型非结合型作为免疫原，分别免疫兔。所得抗血清，抗GLP-1（7-22）血清RO322和抗GLP-1（1-35）血清RO351，用于开发GLP-1区域特异性放射免疫测定（RIA）系统。丙氨酸：由于丙氨酸的c端结构与速激肽相似，因此成功制备了一种针对合成猪丙氨酸（1-15）的单克隆抗体，专门用于免疫组织化学研究。事实上，这种单克隆抗体被发现能够特异性识别产生丙氨酸的细胞，并且不会与任何速激肽发生交叉反应。前LHRH相关肽：已知LHRH前体含有LHRH和一个56个氨基酸的GAP肽。利用合成的GAP（34-56）和proLHRH(1-26)，我们开发了人促lhrh羧基和氨基末端特异性RIA，并生产了抗GAP（34-56）血清（RO242）和抗proLHRH（1-26）血清（RO620）。在家兔中制备了抗合成大鼠GAP(28-51) （RO246）的血清。少

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