Collagen Purification Solutions and Case Sharing

Collagen is a kind of biological macromolecule commonly found in vertebrates. It is abundant in mammalian tissues and organs, usually accounting for 30% of the total protein content of the body. In the human body, collagen widely exists in skin, bones, muscles and other tissues. It mainly plays a role in supporting, repairing and protecting tissue cells. It also participates in physiological and biochemical behaviors such as cell proliferation, differentiation, migration and signal transmission. It is a key structural protein that builds the framework of the body.

Although collagen is one of the most critical proteins in human body, its composition and structure are relatively simple. It is a fibrous protein assembled from multiple procollagen. Procollagen, as the basic unit of collagen, consists of three polypeptide chains, and its primary structure has (Gly-X-Y) n repeats, where X and Y are usually proline and hydroxyproline, and a few are lysine and hydroxylysine. Three procollagen peptide chains coiled around each other via interchain hydrogen bonds to form a stable triple helix structure (Fig. 1). The stability of triple helix conformation of procollagen depends on the strong hydrogen bond formed by hydroxyproline hydroxyl group, which can improve the strength of collagen. The combination of proline and hydroxyproline can make the triple helix structure of collagen twist and twist sharply, and improve the stability of collagen.

Figure 1: Three procollagen peptide chains coil intertwine through interchain hydrogen bonds to form a stable triple helix structure

The source of natural collagen is narrow, and it is mainly extracted by obtaining terrestrial animal collagen (such as pig skin and cow skin) and aquatic animal collagen raw materials (such as fish skin, fish bone and fish scale) by traditional methods such as hot water extraction, acid-base hydrolysis and enzymatic hydrolysis. In addition, due to some religious beliefs of human beings and the risk of zoonotic diseases, people began to look for other safer and more effective collagen preparation methods.

With the rapid development and maturity of genetic engineering, protein engineering and other disciplines, the research of recombinant collagen has also accelerated. Compared with traditional extraction methods, recombinant collagen products have higher purity, stability and safety. With the continuous development of recombinant collagen technology, its market share has also increased year by year (Figure 2).

Figure 2: 2021-2027E China Collagen Market Share

(Image source: Sullivan: 2025 Global Recombinant Type III Collagen Market Research Report)

Analysis of the Production Process of Recombinant Collagen

At present, in the field of upstream fermentation of recombinant collagen, fragment recombinant collagen is mainly prepared by prokaryotic/yeast expression system, while full-length recombinant collagen is mainly produced by mammalian cell expression system (such as CHO cell expression system).

BioLink can provide bioreactors on the scale of 10 L-2000 L, which can meet the preparation needs from lab scale, pilot scale to industrial level. These include single-use rocking bioreactors, single-use stirred-tank bioreactors and other bioreactors, as well as benchtop glass bioreactors commonly used for process development and optimization at laboratory scale.

In the field of downstream purification of recombinant collagen, chromatography has begun to play an increasingly important role in recent years. Theoretically, most collagen has PI ≥ 7. 0, so ion exchange chromatography technology is often used in the purification and preparation of collagen items. Cation exchange chromatography can effectively capture collagen molecules and achieve rapid separation of targets and impurities, while anion exchange chromatography can be used to remove trace impurities such as endotoxin by using flow-through mode. In addition, collagen has good hydrophobicity, so hydrophobic resins are also commonly used resins for collagen projects.

As a biopharmaceutical supplier with mature technology and wide business coverage, BioLink can provide solutions for downstream purification of core equipment and consumables of all scales, including resin, chromatography columns and chromatography systems.

General Technical Route of Recombinant Collagen

Figure 3: BioLink's overall solution for recombinant collagen

Application Case

Case 1: Development of Recombinant Collagen (type III) Fragment Purification Process

Figure 4: (Left) SP Chromstar^® XL one-step purification profile;

(Right) SEC-HPLC test diagram

In this case, the target protein is a 55 kDa recombinant type III collagen fragment, which is secreted and expressed by yeast. The project requires a one-step purification so that SEC-HPLC purity is ≥ 95%. Via SP Chromstar^® XL was purified in one step with a loading of 140 g/L, and the eluted sample SEC-HPLC purity was 98.50%.

Case 2: Development of Recombinant Collagen (type V) Fragment Purification Process

Figure 5: (Left) MaXtar^® MMC HR one-step purification profile; (Right) Gel electrophoresis

In this case, the target protein is expressed by yeast and is a recombinant collagen type V fragment with a size of 26.9 kDa. The project requires one-step purification with purity ≥ 95% and yield ≥ 85%. Via MaXtar^® MMC HR was purified in one step, and the electrophoretic purity of the eluted sample was > 95%, and the yield was 90%.

Case 3: Development of Recombinant Collagen (type I) Fragment Purification Process

Figure 6: (Left) MaXtar^® MMC HR purification profile;

(Right) SEC-HPLC chromatogram

In this case, the target protein is a type Ⅰ recombinant collagen fragment. After on-step MaXtar^® MMC HR, the SEC-HPLC purity of the eluted sample was 95.09% and the yield was 80%.

Introduction to BioLink Suzhou Application Center

Covering an area of 3000sqm, it is equipped with R&D office area, cell culture laboratory, microbiology laboratory, purification laboratory, pilot workshop, and analysis laboratory. Suzhou Application Center is committed to providing customer service with upstream and downstream process development, lab scale and pilot-scale sample preparation, validation and training services.