کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6451902 | 1416986 | 2017 | 4 صفحه PDF | دانلود رایگان |
- Integrating peptidomimetics from phage display biopanning with magnetic particles for immunomagnetic separation of an antigen.
- Identifying functional peptides as a replacement for antibody in immunodiagnostics.
- Potential one-step, low-cost bioseparation of a minute-concentration antigen from a mixture.
- Process metrics for affinity evaluation of peptidomimetics and capture efficiency of immune-magnetics.
Phage display biopanning is a powerful in vitro selection process for screening and identifying peptides that bind to a target protein of interest. With the aim of replacing antibodies in immuno-diagnostic applications, we identified peptides whose binding characteristics mimicked those of anti-human myeloperoxidase (hMPO), a biomarker for acute cardiac diseases. Based on ELISA results from four phage clones, we selected and chemically synthesized a 12-mer peptide (SYIEPPERHRHR). Quartz crystal microbalance and surface plasmon resonance analyses revealed that the molar binding equilibrium ratio of the synthesized peptide was 0.023, approximately 43-fold lower than that of the anti-hMPO antibody. The dissociation constant (Kd) was 57Â nM, which was comparable to that of the native antibody (83Â nM). Next, we biotinylated the peptide at its N-terminus and attached the biotinylated peptide to the surface of streptavidin-coated magnetic particles to assess its ability to selectively capture hMPO. The binding equilibrium data were similar to the previous analyses; specifically, around 0.021Â mol peptide bound to 1Â mol of hMPO. Antigen capture was found to be selective and to be relatively little influenced by the presence of human serum albumin (HSA), an abundant constituent of serum. Our work demonstrates the potential of immunomagnetic isolation to achieve selective capture of a low-concentration antigen from complex solutions such as serum.
Journal: Journal of Biotechnology - Volume 257, 10 September 2017, Pages 118-121