Synthesis and biological characterization of human monocyte chemoattractant protein 1 (MCP-1) and its analogs.
Novel analogs of human monocyte chemoattractant protein 1 (MCP-1) were designed, synthesized and characterized to be used as tools to generate monoclonal antibodies as potential human therapeutics. MCP-1 and three analogs were synthesized by step-wise Fmoc solid phase synthesis. After oxidation to form the two-disulfide bonds, affinity chromatography using an immobilized mouse anti-human MCP-1 monoclonal antibody (mAb) was utilized for a simple and highly effective purification procedure for the proteins. The final products were extensively characterized and compared with recombinant human MCP-1 (rhMCP-1). All proteins showed identical binding with mouse anti-human MCP-1 mAbs as measured by surface plasmon resonance. Synthetic MCP-1 and the analogs were comparable to recombinant MCP-1 in competition radio-ligand binding to CCR2 receptors on THP-1 cells, and MCP-1-induced, calcium mobilization and chemotaxis assays.
Kruszynski
Marian
M
Discovery Research, Centocor, Inc., Radnor, PA, USA. mkruszy2@cntus.jnj.com
Stowell
Nicole
N
Das
Anuk
A
Seideman
Jonathan
J
Tsui
Ping
P
Brigham-Burke
Michael
M
Nemeth
Jennifer F
JF
Sweet
Raymond
R
Heavner
George A
GA
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000494:pharmacology; D001667:Binding, Competitive; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D018932:Chemokine CCL2 / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D002633:Chemotaxis / Q000187:drug effects / Q000502:physiology; D006801:Humans; D008024:Ligands; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011485:Protein Binding; D054390:Receptors, CCR2; D019707:Receptors, Chemokine / Q000187:drug effects; D013329:Structure-Activity Relationship
D000911:Antibodies, Monoclonal; C111835:CCL2 protein, human; C515334:CCR2 protein, human; C515335:Ccr2 protein, mouse; D018932:Chemokine CCL2; D008024:Ligands; D054390:Receptors, CCR2; D019707:Receptors, Chemokine; D002118:Calcium