Chang Z N, Tam M F, Liu C C, Chi C W, Peng H J, Han S H
Faculty of Medical Technology, Research Center for Immunology, National Yang-Ming University, Taipei, Taiwan, Republic of China.
Int Arch Allergy Immunol. 1997 Nov;114(3):258-64. doi: 10.1159/000237677.
Cyn d 1, the group I allergen of Bermuda grass pollen, had been purified and characterized.
A sequential B cell epitope on Cyn d 1 was studied with monoclonal antibodies (MoAbs). Cyn d 1 was cleaved by Achromobacter protease I into fragments, and the resulting peptides were fractionated on reversed-phase columns before being reacted with anti-Cyn d 1 MoAbs in a radioimmunoassay. A Cyn d 1 fragment recognized by its MoAb was selected for Edman degradation. A synthetic peptide was constructed according to the determined sequence.
The epitope on Cyn d 1 recognized by MoAb 18-53 was found to be conformation independent, since its activity was not changed after sodium periodate, guanidine or urea treatment. The enzyme-cleaved fragment containing this epitope was determined to be DVDKPPFDGMTACGNEPIF which corresponds to the N-terminal 46-64 residues of Cyn d 1. The presence of this sequence in the epitope recognized by MoAb 18-53 was demonstrated by enzyme immunoassay and further confirmed by inhibition of binding enzyme immunoassay with synthetic peptides. Some cross-reactivity with the N-terminal 45-63 residues of Lol p 1 was also found.
The primary structure of a sequential epitope on Cyn d 1 was determined, and its activity was confirmed with peptides synthesized according to the determined sequence.
百慕大草花粉的I类变应原Cyn d 1已被纯化和鉴定。
用单克隆抗体（MoAbs）研究Cyn d 1上的一个连续B细胞表位。Cyn d 1被无色杆菌蛋白酶I切割成片段，所得肽段在反相柱上进行分离，然后在放射免疫分析中与抗Cyn d 1 MoAbs反应。选择一个被其MoAb识别的Cyn d 1片段进行埃德曼降解。根据确定的序列合成一个合成肽。
发现单克隆抗体18 - 53识别的Cyn d 1上的表位与构象无关，因为其活性在高碘酸钠、胍或尿素处理后没有改变。确定含有该表位的酶切片段为DVDKPPFDGMTACGNEPIF，对应于Cyn d 1的N端46 - 64个残基。通过酶免疫分析证明了该序列在单克隆抗体18 - 53识别的表位中的存在，并通过合成肽抑制结合酶免疫分析进一步证实。还发现与Lol p 1的N端45 - 63个残基有一些交叉反应性。
确定了Cyn d 1上一个连续表位的一级结构，并用根据确定序列合成的肽证实了其活性。
