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

脑肠激素的合成：我们合成了几种脑肠激素及其相关多肽，含有15-20个氨基酸残基。这些多肽的合成是使用自动合成的Beckman Model 990根据固相技术进行的。在PAM树脂(应用生物系统)上构建了受保护的多肽链。酸性水解物和AP-M酶的氨基酸分析、C&lt；18&Gt；本研究合成了几种新的脑肠激素，如猪和大鼠甘丙素、Helodermin、豚鼠VIP、大鼠PHI、人胰高血糖素样肽-1(GLP-1)、人NPY、人和大鼠促性腺激素释放激素相关肽(GAP)等人和大鼠促性腺激素释放激素相关肽(GAP)。制备抗脑肠激素GLP-1以生产…选择较多的GLP-1特异性抗血清、合成的猪甲状腺球蛋白结合型GLP-1(7-22)和非结合型GLP-1(1-35)作为免疫原，分别免疫兔。制备的抗血清，抗GLP-1(7-22)血清RO322和抗GLP-1(1-35)血清RO351，用于建立GLP-1区域特异性的放射免疫分析(RIA)系统。甘丙素：由于甘丙素在其C末端具有与速激肽相似的结构，因此成功地制备了一种针对合成的猪甘丙素(1-15)的单抗，专门用于免疫组织化学研究。事实上，这种单抗被发现能特异性识别甘丙肽产生细胞，并且不与任何速激肽发生交叉反应。前LHRH相关肽：LHRH前体已知含有LHRH，与56个氨基酸的肽有差距。以合成的GAP(34-56)和proLHRH(1-26)为原料，制备抗GAP(34-56)血清(RO242)和抗ProLHRH(1-26)血清(RO620)，建立人LHRH羧基和氨基末端特异性RIA。合成的大鼠GAP(28-51)(RO246)的抗血清也在兔体内产生。较少

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