The specific binding of peptide ligands to cardiomyocytes derived from mouse embryonic stem cells

Abstract

Purification of pluripotent stem cell (PSC)-derived cardiomyocytes is critical for the application of cardiomyocytes both in clinical and basic research. Finding a specific cell marker is a promising method for purifying induced cells. The present study employed phage display technology to search for particular cell markers that could bind specifically to PSC-derived cardiomyocytes. After three rounds of biopanning, several peptides were obtained. The ELISA results show the no. 3 sequence peptide (QPFTTSLTPPAR), and other four sequences having a consensus motif [SS(Q)PPQ(S)], no. 9, 11, 14, and 10, have relatively high affinity and specificity to cardiomyocytes. Immunofluorescence confirmed that the selected peptides could bind specifically to the PSC-derived cardiomyocytes. Competition tests with chemically synthesized peptides revealed the binding ability was caused by the peptide itself. Western blot analysis proved the phages were both bound to two 17 kDa cardiomyocyte membrane proteins and the no. 9 sequence showed a 55 kDa protein that was not observed in the no. 3 sequence. These results suggest that the selected peptides specifically target receptors on PSC-derived cardiomyocyte membranes. The results will pave the way for further studies of cell surface markers and their applications, such as labeling, purification, and as vehicles for drug delivery.

The development of Army relevant peptide-based surface enhanced Raman scattering (SERS) sensors for biological threat detection

“…The biopanning process typically involves four main steps: preparation of the phage displayed peptide libraries, capture of specific phage that binds to the target, washing of low affinity or non specific phages from the cell surface, and then finally recovery through elution of the enriched target binders for the next round of selection. [74][75][76][77] Using these types of selection techniques, it has been possible to identify and select several different material specific binding peptide sequences (examples include metal compounds, semiconductor materials, minerals, and polymers) 72 as well as particular targets (chemical [78][79][80][81] , biological 65,[82][83] , or energetic hazards [84][85][86][87][88] ). The screening and selection of such phage displayed target binding peptides has attracted particular interest in the research areas of nanotechnology and sensor design.…”

Bacteriophage-based biomaterials for tissue regeneration

“…Han et al[119] successfully used the RGD-phage functionalized PLGA to promote the proliferation and myogenic differentiation of CAC12 myoblasts in the presence of graphene oxide. Ma et al identified a peptide, QPFTTSLTPPAR, which can specifically bind to the mouse ESC-derived cardiomyocytes through phage biopanning using a Ph.D.-12 phage library, providing a cardiomyocytetargeting molecule for further regeneration applications[98]. In vivo phage display derived functional peptides for cardiac regeneration-Robbins et al identified a cardiac targeting peptide, APWHLSSQYSRT, through in vivo phage biopanning using a Ph.D.-12 phage library, providing a cardiachoming molecule to deliver drugs/genes to the heart[99].…”

Targeting biological sensing with commercial SERS substrates

“…The biopanning process typically involves four main steps: preparation of the phage displayed peptide libraries, capture of specific phage that binds to the target, washing of low affinity or non specific phages from the cell surface, and then finally recovery through elution of the enriched target binders for the next round of selection. (25)(26)(27)(28) Using these types of selection techniques, it has been possible to identify and select several different material specific binding peptide sequences (examples include metal compounds, semiconductor materials, minerals, and polymers) (23) as well as particular targets (chemical (29)(30)(31)(32) , biological (16,33,34) , or energetic hazards (35)(36)(37)(38)(39) ). The screening and selection of such phage displayed target binding peptides has attracted particular interest in the research areas of nanotechnology and sensor design.…”