title: Identification of peptides that selectively bind to myoglobin by biopanning of phage displayed-peptide library.
abstract: Biopanning of phage displayed-peptide library was performed against myoglobin, a marker for the early assessment of acute myocardial infarction (AMI), to identify peptides that selectively bind to myoglobin. Using myoglobin-conjugated magnetic beads, phages that bound to myoglobin were collected and amplified for the next round of screening. A 148-fold enrichment of phage titer was observed after five rounds of screening relative to the first round. After phage binding ELISA, three phage clones were selected (3R1, 3R7 and 3R10) and the inserted peptides were chemically synthesized. The analysis of binding affinity showed that the 3R7 (CPSTLGASC) peptide had higher binding affinity (Kd=57 nM) than did the 3R1 (CNLSSSWIC) and 3R10 (CVPRLSAPC) peptide (Kd=125 nM and 293 nM, respectively). Cross binding activity to other proteins, such as bovine serum albumin, troponin I, and creatine kinase-MB, was minimal. In a peptide-antibody sandwich ELISA, the selected peptides efficiently captured myoglobin. Moreover, the concentrations of myoglobin in serum samples measured by a peptide-peptide sandwich assay were comparable to those measured by a commercial antibody-based kit. These results indicate that the identified peptides can be used for the detection of myoglobin and may be a cost effective alternative to antibodies.
authors: Department of Biochemistry and Cell Biology, School of Medicine, Kyungpook National University, Daegu, Republic of Korea; Cell & Matrix Research Institute, Kyungpook National University, Daegu, Republic of Korea; BK21 Plus KNU Biomedical Convergence Program, Department of Biomedical Science, Kyungpook National University, Daegu, Republic of Korea.
authors: Guruprasath
authors: G
authors: Padmanaban
authors: Department of Biochemistry and Cell Biology, School of Medicine, Kyungpook National University, Daegu, Republic of Korea.
authors: Hyekyung
authors: H
authors: Park
authors: Department of Chemical Engineering, Hanyang University, Gyeonggi-do, Republic of Korea.
authors: Ji Suk
authors: JS
authors: Choi
authors: Department of Chemical Engineering, Hanyang University, Gyeonggi-do, Republic of Korea.
authors: Yong-Woo
authors: YW
authors: Cho
authors: Department of Cardiology, Gil Medical Center, Gachon University of Medicine and Science, Incheon, Republic of Korea.
authors: Woong Chol
authors: WC
authors: Kang
authors: Department of Cardiology, School of Medicine, Hanyang University, Seoul, Republic of Korea.
authors: Chan-Il
authors: CI
authors: Moon
authors: Biomedical Research Institute, KIST, Seoul, Republic of Korea.
authors: In-San
authors: IS
authors: Kim
authors: Department of Biochemistry and Cell Biology, School of Medicine, Kyungpook National University, Daegu, Republic of Korea; Cell & Matrix Research Institute, Kyungpook National University, Daegu, Republic of Korea; BK21 Plus KNU Biomedical Convergence Program, Department of Biomedical Science, Kyungpook National University, Daegu, Republic of Korea. Electronic address: leebh@knu.ac.kr.
authors: Byung-Heon
authors: BH
authors: Lee
mesh_terms: D015415:Biomarkers / Q000097:blood* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006331:Heart Diseases / Q000097:blood; D006801:Humans; D009211:Myoglobin / Q000097:blood* / Q000378:metabolism; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding; D015203:Reproducibility of Results
chemical_list: D015415:Biomarkers; D009211:Myoglobin; D019151:Peptide Library; D010455:Peptides
keywords: Cardiac biomarkers; Myoglobin; Peptide; Phage display; Sandwich assays