De novo assisted AFB1-Specific monoclonal antibody sequence assembly and comprehensive molecular characterization.

Despite their widely used and access as biological reagents in analytical methods, the detailed structural features for most of the antibodies were rarely known. Here, a new antibody for AFB1 with high specificity in constructing ELISA was studied in detail. The molecular structure and modification were elucidated mainly by nano-electrospray ionization mass spectrometry. The mass experiments, including MALDI-TOF MS, revealed complete and specific fragments, including antibody molecular weight, peptides, glycopeptide, and N-glycoform. By proteolytic treatment of pepsin and trypsin and high-resolution tandem-MS, the primary structure of the newly developed anti-AFB1 antibody was assembled by several rounds of Database search process assisted with the de novo results. The antibody CDR annotation and constraint-based multiple alignment tool were used to differentiate and align the sequences. The method uses only two proteases to generate numerous peptides for de novo sequencing. This artificial assembled AFB1-specific monoclonal antibody sequence was validated by comparison with the sequencing results of the immunoglobulin gene. The results showed that this method achieves full sequence coverage of anti-AFB1 monoclonal antibody, with an accuracy of 100% in the CDR regions of light chain and four amino acid mismatch in heavy chain. This simple and low-cost method was confirmed by treating a public dataset. The secondary structure information of intact antibody was also elucidated from the results of circular dichroism spectrum.

Effects of food components and processing parameters on plant-based meat texture formation and evaluation methods.

Meat is the main source of protein nutrition for humans. However, given the increasing population and environmental pressure, a shortage of meat is expected in the future. The preparation of plant-based (meat) products using existing processing methods represents a breakthrough to solve the shortage of traditional animal meat in the future, and texture is a key issue determining whether plant-based (meat) products can replace traditional animal meat. Therefore, this review highlights recent progress in the texture of plant-based (meat) products. First, the texture changes in plant-based (meat) products were reviewed based on components (proteins, starch, lipids, food additives) and processing methods (processing factors, instrumental factors). Then, the current methods used to analyze the texture of plant-based (meat) products were summarized based on sensory evaluation and instrumental analysis (mechanical analysis, spectral analysis, and image analysis). Finally, this review outlined methods to improve the texture quality of plant-based (meat) products and analytical evaluation methods and provided an outlook on future research priorities.