Scientific Publications by FDA Staff

Rapid Commun Mass Spectrom 2020 Apr 15;34(7):e8639

Proteomics profiling of swine serum following lipopolysaccharide stimulation.

Olumee-Shabon Z, Chattopadhaya C, Myers MJ

RATIONALE: There are no approved animal drugs for management of inflammation in swine due to lack of validated animal models. To assess efficacy, it was essential to develop proteomics approaches to identify suitable biomarkers of inflammation as presented in this study. METHODS: Serum samples were collected from a group of 4 pigs prior to (baseline), and 24 and 48 h following lipopolysaccharide (LPS) stimulation to reveal proteomic changes during inflammation. Two other pigs served as untreated controls. Proteins were separated by either 1D SDS-PAGE or 2D gel electrophoresis (2DE) prior to analysis by nano flow liquid chromatography (nLC) coupled to tandem mass spectrometry. RESULTS: We identified 165 proteins using SDS-PAGE, of which 47 proteins were also detected by 2DE prior to nLC-MS/MS. More than half (72%) of all characterized proteins were modulated as a result of LPS stimulation, many of which are known to be involved with innate and adaptive immunity. Pig serum samples obtained 24 h after LPS initiation of inflammation showed protein modulations of serum albumin, serotransferrin, light and heavy immunoglobulin chains (IGs), and major acute phase proteins including haptoglobin (HPT), serum amyloid A2 (SAA2), C-reactive protein (CRP), beta-2-glycoprotein 1 (B-2GP1), alpha-2-HS-glycoprotein (A2HS), alpha-1-antitrypsin (A1AT), and alpha-1-acid glycoprotein (A1AG). SAA2 was distinguished from the other SAA isoforms by the unique peptide sequence of SAA2. CONCLUSIONS: The results provided proteomic analysis of swine serum due to LPS stimulation and indicated the importance of SAA2, which appears to be unique and may be regarded as a potential clinical diagnostic biomarker of inflammation.