Production of recombinant collagen: state of the art and challenges

Abstract

Collagen, which is often used in healthcare materials and biomedical research, is largely extracted from animal sources. Recombinant human collagen has the potential to be a promising alternative to animal collagen which has many shortcomings, including immunogenicity and lack of biocompatibility. Currently, recombinant human collagen has been expressed in both eukaryotic and prokaryotic hosts with varying degrees of success. One issue with recombinant collagen across all hosts is the inability to achieve full length collagen with native amounts of post-translational modifications, prompting much exciting research in this direction. There has also been much effort in improving yield and biomimicry of recombinant collagen. This review discusses collagen structure and current methods for extracting animal collagen, before introducing current research in synthesising recombinant human collagen in various hosts, and finally highlighting challenges in the field.

Additive Biomanufacturing with Collagen Inks

“…Cells such as fibroblast and chondrocytes which specialize in type I and type II collagen production respectively can be cultured and the synthesised collagen harvested from media or cell layers. Recombinant collagen production is using genetically engineered microorganisms, plants or animals such as bacteria, yeast, transgenic corn and silkworms [29,30]. Synthetic peptides mimicking collagen trimeric structure have also been investigated to produce collagen-like peptides [31,32].…”

Plant molecular farming: production of metallic nanoparticles and therapeutic proteins using green factories

“…148 Pro-collagen (commercial collagen) is prepared from domesticated animals, including pigs and cows and has an increasing global demand. 149,150 Inauspiciously, it is liable to harbor human pathogens, comprising prions or viruses. 149 As a viable alternative, tobacco plants have been used to efficiently express human recombinant type I pro-collagen using transgenic technology.…”

Method for purification of collagen: A systematic review

“…This is due to the outbreak of transmissible diseases like bovine spongiform encephalopathy (BSE), transmissible spongiform encephalopathies (TSE) and foot-and-mouth disease (FMD) that pose threat to human (Jongjareonrak et al, 2005;Peng et al, 2012a;Silvipriya et al, 2015;Singh et al, 2011;Swatschek et al, 2002;Zhang et al, 2009). Furthermore, religious sentiments for Muslims, Jewish, and Hindus created another issue in the consumption of porcine or bovine-derived collagen (Sadowska et al, 2003;Singh et al, 2011;Souza, 2016;Wang et al, 2017). Islamic and Jewish believers are prohibited in the consumption of pork, whereas bovine sources are prohibited for Sikhs and Hindus (Eriksson et al, 2013;Herbert et al, 1994;Singh et al, 2011;Souza, 2016).…”

“…Purification of recombinant bacterial collagen using a combination of precipitation and proteolysis by pepsin was performed in two out of three studies (Cao & Xu, 2008;Werkmeister & Ramshaw, 2015). Werkmeister, Singh, and coworkers reported that the application of precipitation and proteolysis is applicable to a wide range of bacterial collagen with different collagen domain sizes (Singh et al, 2011;Wang et al, 2017). The acid precipitation method can be considered as an efficient technique in removing a high amount of bacterial host proteins and they will tend to precipitate upon the addition of acid as host proteins were least soluble at a pH lower than 5 (Kiew & Don, 2013;Werkmeister & Ramshaw, 2015).…”

“…The conventional methods consist of several unit operations that lead to a high cost of operation and maintenance (Przybycien et al, 2004;Raja et al, 2012;Song et al, 2013). For instance, precipitation needs to be combined with other processes such as chromatography processes, which are complex, tedious, time-consuming and often produced low yields (Cao & Xu, 2008;Sato et al, 2003;Wang et al, 2017). Furthermore, the high number of unit operations involved causes a high loss of target molecules that will result in low product yield (Xing et al, 2012).…”