Design and Production of collagen-like engineered peptide (CLP) for tissue-engineering applications

Transplantation of biomaterials into patients suffering from organ failure or damage can be conducted instead of receiving grafts from organ donors or animals. Similar to donated grafts, biomaterials aim at replacing the functions of lost organs, and saving patients’ life or improving their quality of life. Hopefully, a potential alternative to conventional biomaterials are synthetic peptides that can be constructed easily in the laboratory, be processed easily, and have a controlled degradation pattern. Collagen-like peptides (CLP), also known as collagen mimetic peptides, are relatively short sequences that have been designed to replicate and reproduce the function of full-length collagen and developed as functional alternatives of collagen. This study aims at producing CLP proteins that later were used for hydrogel formation. The clp nucleotide sequences cloned into pColdI vectors and transformed into ClearColi bacteria for protein expression. Restriction enzyme analyses followed by agarose gel electrophoreses as well as sequencing analyses were performed to verify the integrity of the constructs. Further, the conditions for the proteins’ expression were optimized and a strategy for the isolation and purification of proteins were developed by western blotting and Ni-NTA column chromatography. Further, CLP peptides were used to make collagen hydrogels as scaffolds for 3D cell culture. The hydrogels prove to be suitable scaffolds for the generation of artificial tissues that could serve as transplants and improve the lives of many patients that suffer from organ failure or damage.