1. Evaluation of GC-MS for identification and characterization of pneumococcal serotype 24A, 24B, and 24F capsular polysaccharide.
- Author
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Shende, Niraj, Karale, Abhijeet, Deshpande, Hrishikesh, Belapurkar, Hrushikesh, Gulhane, Ashish, Bhagade, Sudhakar, Bore, Prashant, Soni, Dipen, Marathe, Preeti, Patni, Sushil, Dhere, Rajeev, and Mallya, Asha
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GAS chromatography/Mass spectrometry (GC-MS) , *STEREOISOMERS , *CHROMATOGRAMS , *IMMUNE response , *SEROTYPES - Abstract
Analysis of pneumococcal polysaccharides (PnPs) has been an arduous task, especially in similar serotypes. Pneumococci invades the host immune response by modulating capsule structure with small genetic changes making them indistinguishable from similar serotypes by conventional modes of analysis. The new serotype 24F causing invasive pneumococcal-resistant infection is an analytical challenge for its analysis as related serotypes 24A and 24B Ps share a common backbone. The difference in the branched chain which contains arabinitol and ribitol in 24F and 24B respectively are stereoisomers making their identification even more challenging. The composition analysis by GC-MS revealed distinct peaks for arabinitol in 24F and 24A Ps and ribitol in Pn 24B serotype polysaccharide. The mass spectral analysis confirmed their identification along with a heterologous cross-reactivity which confirmed anti-Pn-24F mAb reactive to Pn 24B than Pn 24A. The quantitative analysis of pneumococcal 24A, 24B and 24F using GC-MS showed sensitive analysis over the concentration range 3.125–200 μg/mL with regression coefficient >0.99 making ideal modality for the characterization, identification, and quantitation of pneumococcal 24A, 24B and 24F similar serotypes. [Display omitted] • GC-MS separates the stereoisomeric sugars present in pneumococcal 24 serotypes. • GC-MS provides the unique total ion chromatogram to distinguish pneumococcal 24A, 24B and 24F serotype. • Mass spectral analysis of Pn 24 serotype provides unique identification of arabinitol and ribitol stereoisomers. • Heterologous cross-reactivity of pneumococcal 24F mAb was observed with 24B polysaccharide by bead-based assay. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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