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Development of Single‐Chain Antibodies Specific to Lysophosphatidic Acid Receptor 2
Author(s) -
Son Sang Hyeon,
Baek Seungil,
Ju ManSeok,
Han SeongGu,
Jung Sang Taek,
Yu Yeon Gyu
Publication year - 2018
Publication title -
bulletin of the korean chemical society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.237
H-Index - 59
ISSN - 1229-5949
DOI - 10.1002/bkcs.11414
Subject(s) - lysophosphatidic acid , receptor , antibody , recombinant dna , microbiology and biotechnology , phage display , biopanning , biology , cell growth , complementarity determining region , monoclonal antibody , chemistry , biochemistry , peptide library , peptide sequence , immunology , gene
Lysophosphatidic acids (LPAs) are ubiquitous serum phospholipids that trigger diverse cellular responses such as cell proliferation, migration, cell survival, and calcium influx. LPA receptors belong to the G‐protein coupled receptor (GPCR) family, and seven subtypes of LPA receptors have been identified. Among them, LPA receptor 2 (LPA 2 ) is involved in the proliferation and metastasis of ovarian, cervical, and breast cancers. Hence, LPA 2 ‐specific antagonists or antibodies are considered as potential anticancer therapeutics. To develop antibodies against LPA 2 , a recombinant LPA 2 was expressed in Escherichia coli , purified to homogeneity, and immobilized on a solid surface as an active conformation. An M13 phage library displaying single‐chain variable fragments (scFvs) of human IgG containing randomized complementarity‐determining regions was applied to select LPA 2 ‐specific scFv clones. After five rounds of biopanning, a few scFv clones which showed specific binding to LPA 2 were identified. Single‐chain antibodies (scAbs), which contain the isolated scFvs and Cκ domain, were constructed and expressed in E. coli . The purified scAbs showed specific binding to LPA 2 with K D values of 200–600 nM.