Tyutyulkova S, Paul S
Department of Anesthesiology, University of Nebraska Medical Center, Omaha.
Appl Biochem Biotechnol. 1994 May-Jun;47(2-3):191-7; discussion 198. doi: 10.1007/BF02787934.
Human kappa-light chains (L chains) were amplified by the reverse transcriptase-polymerase chain reaction (PCR) and cloned into a phagemid vector. Phage particles displaying L chains were fractionated on immobilized vasoactive intestinal peptide (VIP). The resultant phage preparation displayed saturable binding of (tyr10-125I)VIP. One of the L-chain clones (hk13) was deduced to be related to subgroup I of kappa-light chains based on its nucleotide sequence. The VIP binding activity of the soluble and phage-displayed form of this L chain was confirmed by radioimmunoassay and ELISA, respectively. These observations demonstrate the potential of selecting antigen-specific L chains from phage-display libraries.
人κ轻链（L链）通过逆转录聚合酶链反应（PCR）进行扩增，并克隆到噬菌粒载体中。展示L链的噬菌体颗粒在固定化血管活性肠肽（VIP）上进行分级分离。所得噬菌体制剂显示出（tyr10-125I）VIP的饱和结合。根据其核苷酸序列，其中一个L链克隆（hk13）被推断与κ轻链的I亚组相关。分别通过放射免疫测定和酶联免疫吸附测定证实了该L链的可溶性形式和噬菌体展示形式的VIP结合活性。这些观察结果证明了从噬菌体展示文库中选择抗原特异性L链的潜力。
