Screening and validation of an alkaline-tolerant biomimetic affinity chromatography A5-87 resin for purification of discarded bovine serum Immunoglobulin G.

It is important to recycle the bovine blood discarded at slaughter and develop it into high value-added bovine serum products. Biomimetic affinity chromatography (BiAC) resins have been developed to specifically purify bovine serum immunoglobulin G (Bs-IgG). The BiAC strategy was used to screen the resins with the best purification effect on Bs-IgG. Four resins with specificity for Bs-IgG adsorption were selected from 90 BiAC resins. Finally, BiAC-A5-87 was selected and used to purify Bs-IgG based on the results of SDS-PAGE and BCA protein quantification analysis. The adsorption capacity and purity of BiAC-A5-87 were 32.79 ± 3.57 mg/mL and 85.9 ± 1.21 % for Bs-IgG, respectively. The total protein recovery rate of Bs-IgG purified by BiAC-A5-87 was 89.78±3.52 %. The resin of BiAC-A5-87 column was recycled in 40 breakthrough cycles, and its Bs-IgG adsorption efficiency decreased by less than 10 %. After soaking BiAC-A5-87 in 1.0 moL NaOH solution for 64 h, its adsorption capacity for Bs-IgG was almost the same as that before soaking. The development of waste bovine serum not only realizes the utilization of blood resources and produces high economic benefits but also reduces the pollution of the environment.

Improving the profile and reliability of cytoplasmic proteins from M. tuberculosis using biomimetic affinity chromatography coupled with LC-MS/MS analysis.

M. tuberculosis is the most successful intracellular pathogen and remains a major threat to human health. It is crucial to investigate the profile of cytoplasmic proteins from M. tuberculosis for pathogenesis, clinical markers, and protein vaccine development. In this study, six biomimetic affinity chromatography (BiAC) resins with high differences were selected for M. tuberculosis-cytoplasmic protein fractionation. All fractions were identified using liquid chromatography-mass spectrometry (LC-MS/MS) analysis. A total of 1246 M. tuberculosis proteins were detected (p < 0.05), among which 1092 M. tuberculosis proteins were identified in BiAC fractionations and 714 M. tuberculosis proteins in un-fractionations (Table S1.3.1). The majority of 66.8% (831/1246) identifications were distributed in the range of Mw 7.0-70.0 kDa, pI 3.5-8.0, and Gravy values <0.3. Furthermore, 560 M. tuberculosis proteins were detected in both the BiAC fractionations and un-fractionations. Compared with the un-fractionations, the average number of prot_matches, prot_cover, prot_sequence, and emPAI values of these 560 proteins in the BiAC fractionations were increased by 3.791, 1.420, 1.307, and 1.788 times, respectively. Overall, compared with un-fractionations, the confidence and profile of M. tuberculosis cytoplasmic proteins were improved by BiAC fractionations coupled with LC-MS/MS analysis. The strategy of BiAC fractionation can be used as an effective method for pre-separating protein mixtures in proteomic studies.

Predicted molecules and signaling pathways for regulating seizures in the hippocampus in lithium-pilocarpine induced acute epileptic rats: A proteomics study.

Seizures in rodent models that are induced by lithium-pilocarpine mimic human seizures in a highly isomorphic manner. The hippocampus is a brain region that generates and spreads seizures. In order to understand the early phases of seizure events occurring in the hippocampus, global protein expression levels in the hippocampus on day 1 and day 3 were analyzed in lithium-pilocarpine induced acute epileptic rat models using a tandem mass tag-based proteomic approach. Our results showed that differentially expressed proteins were likely to be enhanced rather than prohibited in modulating seizure activity on days 1 and 3 in lithium-pilocarpine induced seizure rats. The differentially regulated proteins differed on days 1 and 3 in the seizure rats, indicating that different molecules and pathways are involved in seizure events occurring from day 1 to day 3 following lithium-pilocarpine administration. In regard to subcellular distribution, the results suggest that post-seizure cellular function in the hippocampus is possibly regulated in a differential manner on seizure progression. Gene ontology annotation results showed that, on day 1 following lithium-pilocarpine administration, it is likely necessary to regulate macromolecular complex assembly, and cell death, while on day 3, it may be necessary to modulate protein metabolic process, cytoplasm, and protein binding. Protein metabolic process rather than macromolecular complex assembly and cell death were affected on day 3 following lithium-pilocarpine administration. The extracellular matrix, receptors, and the constitution of plasma membranes were altered most strongly in the development of seizure events. In a KEGG pathway enrichment cluster analysis, the signaling pathways identified were relevant to sustained angiogenesis and evading apoptosis, and complement and coagulation cascades. On day 3, pathways relevant to Huntington's disease, and tumor necrosis factor signaling were most prevalent. These results suggest that seizure events occurring in day 1 modulate macromolecular complex assembly and cell death, and in day 3 modulate biological protein metabolic process. In summary, our study found limited evidence for ongoing seizure events in the hippocampus of lithium-pilocarpine induced animal models; nevertheless, evaluating the global differential expression of proteins and their impacts on bio-function may offer new perspectives for studying epileptogenesis in the future.