Chung Junho, Park Sojung, Kim Dongjo, Rhim Junghyo, Kim Ikjung, Choi Inhak, Yi Kyesook, Ryu Sungho, Suh Pannghill, Chung Doohyun, Bae Youngmee, Shin Youngkee, Park Sunghoe
National Cancer Center, Ilsan-gu, Goyang, Gyeonggi, Korea.
J Cancer Res Clin Oncol. 2002 Dec;128(12):641-9. doi: 10.1007/s00432-002-0390-x. Epub 2002 Nov 13.
In the present study an antigen-mimetic peptide of the anti-JL1 leukemia-specific monoclonal antibody (mAb) was identified and characterized.
From combinatorial peptide phage display libraries displaying the random linear heptapeptides and dodecapeptides, we selected clones with affinity to anti-JL1 mAb through repeated rounds of panning on a mAb-coated ELISA plate. The antigenicity and immunogenicity of the peptide epitopes were then studied using chemically synthesized peptides.
The selected clones had the LXPSIP consensus sequence. Two synthetic peptides LPPSIPFGLTVGGGGS and LLPSIPNQAYLGGGGS specifically reacted with anti-JL1 mAb in ELISA. These two peptides were found to inhibit the interaction between anti-JL1 mAb and JL1 antigen-positive Molt-4 cells. Although the immune sera raised against the keyhole limpet hemocyanin-conjugated peptides failed to react with Molt-4 cells, it showed strong reactivity to the peptide epitope. However, one mAb raised by peptide immunization successfully bound to Molt-4 cells.
An epitope-mimetic peptide of anti-JL1 mAb was found using combinatorial peptide phage display libraries. It induced strong humoral response against itself, but only a limited fraction of this humoral response was cross-reactive with the original JL1 antigen.
在本研究中，鉴定并表征了抗JL1白血病特异性单克隆抗体（mAb）的一种抗原模拟肽。
从展示随机线性七肽和十二肽的组合肽噬菌体展示文库中，通过在包被有mAb的ELISA板上进行多轮淘选，我们筛选出了与抗JL1 mAb具有亲和力的克隆。然后使用化学合成肽研究肽表位的抗原性和免疫原性。
筛选出的克隆具有LXPSIP共有序列。两种合成肽LPPSIPFGLTVGGGGS和LLPSIPNQAYLGGGGS在ELISA中与抗JL1 mAb特异性反应。发现这两种肽可抑制抗JL1 mAb与JL1抗原阳性的Molt-4细胞之间的相互作用。尽管针对钥孔戚血蓝蛋白偶联肽产生的免疫血清未能与Molt-4细胞反应，但它对肽表位显示出强反应性。然而，通过肽免疫产生的一种mAb成功地与Molt-4细胞结合。
利用组合肽噬菌体展示文库发现了抗JL1 mAb的一种表位模拟肽。它诱导了针对自身的强烈体液反应，但这种体液反应中只有有限一部分与原始的JL1抗原发生交叉反应。
