Study on Detection Method of Bacterial Endotoxin in Recombinant Collagen and Application of Data Analysis Technology

As an important biomedical material, the safety and quality control of recombinant collagen are particularly important. As a key safety index, bacterial endotoxin may cause serious harm to biological products if it exists in trace amounts. In view of this, this study combined protease degradation and gel method to eliminate matrix effect and optimize the detection process. High purity (<tex xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">$&gt; 98\%$</tex>) recombinant human collagen Type I samples were used in the experiment, and the concentration gradient was established by using Lonza PyroStar ES-F bacterial endotoxin standard. After papain degradation, the solubility and molecular weight of the sample decreased significantly, while the degraded collagen still retained its biocompatibility and low immunogenicity. The content of bacterial endotoxin in the samples before and after degradation was detected by gel method with Lonza PyroGene LAL kit. The results showed that the endotoxin content in all samples was far below the safe limit (<tex xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">$0.25 \text{EU} / \text{ml}$</tex>), indicating that degradation treatment did not increase the endotoxin content. The application of data analysis technology further reveals the significant influence of reaction time and temperature on detection sensitivity. The absorbance value was converted into endotoxin concentration by GraphPad Prism software, and the accuracy and reliability of the detection method were evaluated. ANOVA analysis showed that the treatment conditions had a significant effect on the endotoxin concentration, suggesting that optimizing the treatment conditions could effectively reduce the endotoxin content. Compared with the existing methods, the gel method used in this study showed higher sensitivity (<tex xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">$0.001 \text{EU} / \text{ml}$</tex>), specificity (0.99), simple operation (4.5/5) and lower cost (2/5). This study provides an accurate, sensitive and economical method for the detection of bacterial endotoxin in biological products such as recombinant collagen, and provides a scientific basis for the quality control of biological products.