The identification of affinity peptide ligands specific to the variable region of human antibodies.
Of all potential biological therapeutics, monoclonal antibody (mAb)-based therapies are becoming the dominant focus of clinical research. In particular, smaller recombinant antibody fragments such as single-chain variable fragments (scFv) have become the subject of intense focus. However, an efficient affinity ligand for antibody fragment purification has not been developed. In the present study, we designed a consensus sequence for the human antibody heavy or light chain-variable regions (Fv) based on the antibody sequences available in the ImMunoGeneTics information system (IMGT), and synthesized these consensus sequences as template Fv antibodies. We then screened peptide ligands that specifically bind to the repertoire-derived human Fv consensus antibody using a 12-mer-peptide library expressed-phage display method. Subsequently, 1 peptide for the VH template and 8 peptides for the VK template were selected as the candidate ligands after 4 rounds of panning the phage display. Using peptide-bead-based immunoprecipitation, the code-4 and code-13 peptides showed recovery rates of the VH and VK templates that were 20-30% and 40-50%, respectively. Both peptides exhibited better recovery rates for trastuzumab scFv (approximately 40%). If it were possible to identify the best combination of VH and VK-binding peptides among the ligand peptides suitable for the human mAb Fv sequence, the result could be a promising purification tool that might greatly improve the cost efficiencies of the purification process.
Immunotherapy Division, Shizuoka Cancer Center Research Institute, 1007 Shimonagakubo, Nagaizumicho,Sunto-gun, Shizuoka 411-8777, Japan.
Yasuto
Y
Akiyama
Haruo
H
Miyata
Masaru
M
Komiyama
Masahiro
M
Nogami
Kazumichi
K
Ozawa
Chie
C
Oshita
Akiko
A
Kume
Tadashi
T
Ashizawa
Naoki
N
Sakura
Tohru
T
Mochizuki
Ken
K
Yamaguchi
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000235:genetics*; D019295:Computational Biology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000737:chemistry / Q000235:genetics*; D047468:Immunoprecipitation; D008024:Ligands; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding / Q000276:immunology; D011993:Recombinant Fusion Proteins; D016415:Sequence Alignment; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000235:genetics
D000911:Antibodies, Monoclonal; D007135:Immunoglobulin Variable Region; D008024:Ligands; D019151:Peptide Library; D010455:Peptides; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies