Structural Peculiarities and Biological Properties of the Lipopolysaccharide from Herbaspirillum seropedicae Z78

Velichko, Natalya S.; Surkina, A. K.; Fedonenko, Yu. P.; Zdorovenko, Evelina L.; Коннова, С. А.

doi:10.1134/s002626171805017x

Cited by 6 publications

(8 citation statements)

References 25 publications

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“…Efforts to raise antibodies against purified LPS were unsuccessful, possibly owing to the structural peculiarities of H. seropedicae Z78 LPS. Previous work by us (Velichko et al 2018) has found that the OPSrepeating unit in H. seropedicae Z78 consists of glycerol-1-phosphate substituted by residues of N-acetyl-D-glucosamine. Structures of this kind are typical of the teichoic acids of gram-positive bacteria (Naumova et al 2001) and are rare in gram-negative bacteria (Kondakova et al 2005;Zdorovenko et al 2011;Shashkov et al 2015).…”

Section: Discussionmentioning

confidence: 86%

“…2 ELISA of CPS-I, CPS-II, EPS-I, EPS-II, and LPS by using miniAbs CPS-II prepared after the first (a) and fourth (b) selection rounds Smol´kina et al 2012], the removal of the capsules from the cell surface was absolutely necessary to prevent contamination of the LPS preparations with the surface glycopolymers. The monosaccharide composition of H. seropedicae Z78 CPS-I, CPS-II, EPS-I, and EPS-II was described by us earlier [Smol´kina et al 2012], as were the monosaccharide composition and the structure of H. seropedicae Z78 LPS [Velichko et al 2018]. In this study, we used the following preparations: EPS-I, EPS-II, CPS-I, CPS-II, and LPS.…”

Section: Discussionmentioning

confidence: 96%

“…All carbohydrate-containing fractions that did not give absorption between 240 and 260 nm were pooled, concentrated, and lyophilized. LPS was extracted from the acetone-treated cells (10 g) with hot 45% aqueous phenol by a modified Westphal procedure (Velichko et al 2018), purified by ultracentrifugation two times (each at 105000×g for 4 h), and lyophilized with a Benchtop 2K apparatus (VirTis, USA).…”

Section: Isolation and Purification Of Bacterial Polysaccharidesmentioning

confidence: 99%

“…Purified preparations of CPS-I, CPS-II, EPS-I, EPS-II and LPS were isolated as described by Smol'kina et al (2012) and Velichko et. al.…”

Section: Preparation Of Rabbit Polyclonal Antibodiesmentioning

confidence: 99%

“…We examined the inhibition of miniAbs by commercial rhamnose, galactose, glucose, and N-acetyl-D-glucosamine (all sugars part of LPS), as well as by the OPS and core oligosaccharide of H. seropedicae Z78 (Velichko et al 2018). The negative control was commercial altrose.…”

Section: Inhibition Of the Glycopolymer-miniab Interactionmentioning

confidence: 99%

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Phage antibodies for the immunochemical characterization of Herbaspirillum seropedicae Z78 glycopolymers

Velichko

Fedonenko

2019

Ann Microbiol

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Purpose Microbial carbohydrate antigens are targets of the immune systems of hosts. In this context, it is of interest to obtain data that will permit judgment of the degree of heterogeneity, chemical makeup, and localization of the antigenic determinants of the Herbaspirillum surface glycopolymers. Methods A sheep single-chain antibody-fragment phage library (Griffin.1, UK) was used to obtain miniantibodies to the exopolysaccharides (EPS-I and EPS-II), capsular polysaccharides (CPS-I and CPS-II) and lipopolysaccharide (LPS) of Herbaspirillum seropedicae Z78. To infer about the presence or absence of common antigenic determinants in the cell-surface polysaccharides of H. seropedicae Z78, we ran a comparative immunoassay using rabbit polyclonal and phage recombinant antibodies to the surface glycopolymers of H. seropedicae Z78. Results We isolated and purified the exopolysaccharides (EPS-I and EPS-II), capsular polysaccharides (CPS-I and CPS-II), and lipopolysaccharide (LPS) of Herbaspirillum seropedicae Z78. Using rabbit polyclonal antibodies, we found that these cellsurface polysaccharides were of a complex nature. EPS-I, EPS-II, CPS-I, CPS-II, and LPS contained common antigenic determinants. CPS-I, CPS-II, and LPS also contained individual antigenic determinants composed of rhamnose, N-acetyl-D-glucosamine, and N-acetyl-D-galactosamine-sugars responsible for cross-reactions with miniantibodies. Conclusions The anti-LPS miniantibodies were more specific for the core region of the LPS, in which rhamnose was the most abundant sugar, than they were specific for its O portion. The miniantibodies we isolated can be useful reagents not only in basic biochemical research but also in clinical diagnostic and therapeutic applications.

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