Synthesis and immunoreactivity of neoglycoproteins containing the trisaccharide unit of phenolic glycolipid I of Mycobacterium leprae

Delphi Chatterjee, Sang Nae Cho, Carol Stewart, James T. Douglas, Tsuyoshi Fujiwara, Patrick J. Brennan

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

The trisaccharide segment, O-(3,6-di-O-methyl-β-d-glucopyranosyl)-(1→4)-O-(2,3-di-O-methyl-α-l-rhamnopyranosyl)-(1→2)- 3-O-methyl-l-rhamnopyranose, of the Mycobacterium leprae-specific phenolic glycolipid I has been synthesized as its 8-(methoxycarbonyl)octyl glycoside and coupled to a carrier protein, to produce a leprosy-specific neoglycoprotein, the so-called natural trisaccharide-octyl-bovine serum albumin (NT-O-BSA). Special features of the synthetic strategy were the use of silver trifluoromethanesulfonate (triflate) to promote glycosylation, resulting in the rhamnobiose in high yield and absolute stereospecificity. The terminal 3,6-di-O-methyl-d-glucopyranosyl group was introduced after O-deallylation of the rhamnobiose. Removal of protecting groups yielded the trisaccharide hapten suitable for coupling to carrier protein. Poly(acrylamide)-gel electrophoresis of the neoglycoprotein demonstrated its purity, and subsequent immunoblotting with a monoclonal antibody directed to the terminal 3,6-di-O-methyl-β-d-glucopyranosyl epitope of the native glycolipid demonstrated its antigenicity. Comparative serological testing in enzyme-linked immunosorbent assays of NT-O-BSA, the corresponding disaccharide-containing products, and another trisaccharide-containing neoglycoprotein, O-(3,6-di-O-methyl-β-d-glucopyranosyl)-(1→4)-O-(2,3-di-O-methyl-α-l-rhamnopyranosyl)-(1→2)-(3-O- methyl-α-l-rhamnopyranosyl)-(1→4′)-oxy-(3-phenylpropanoyl)-BSA (NT-P-BSA) [Fujiwara et al., Agric. Biol. Chem., 51 (1987) 2539-2547] against sera from leprosy patients and control populations showed concordance; the presence of the innermost sugar did not contribute significantly to sensitivity or specificity. The di- and tri-saccharide-containing neoantigens, on account of ready availability and solubility, provide greater flexibility than the native glycolipid for the serodiagnosis of leprosy.

Original languageEnglish
Pages (from-to)241-260
Number of pages20
JournalCarbohydrate Research
Volume183
Issue number2
DOIs
Publication statusPublished - 1988 Dec 1

Fingerprint

Trisaccharides
Glycoproteins
Glycolipids
Leprosy
Bovine Serum Albumin
Carrier Proteins
Glycosylation
Immunosorbents
Acrylamide
Haptens
Disaccharides
Serologic Tests
Glycosides
Polyacrylates
Electrophoresis
Immunoblotting
Sugars
Solubility
Epitopes
Assays

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

Cite this

Chatterjee, Delphi ; Cho, Sang Nae ; Stewart, Carol ; Douglas, James T. ; Fujiwara, Tsuyoshi ; Brennan, Patrick J. / Synthesis and immunoreactivity of neoglycoproteins containing the trisaccharide unit of phenolic glycolipid I of Mycobacterium leprae. In: Carbohydrate Research. 1988 ; Vol. 183, No. 2. pp. 241-260.
@article{72ec36f9019143d3a960999bc4eb776c,
title = "Synthesis and immunoreactivity of neoglycoproteins containing the trisaccharide unit of phenolic glycolipid I of Mycobacterium leprae",
abstract = "The trisaccharide segment, O-(3,6-di-O-methyl-β-d-glucopyranosyl)-(1→4)-O-(2,3-di-O-methyl-α-l-rhamnopyranosyl)-(1→2)- 3-O-methyl-l-rhamnopyranose, of the Mycobacterium leprae-specific phenolic glycolipid I has been synthesized as its 8-(methoxycarbonyl)octyl glycoside and coupled to a carrier protein, to produce a leprosy-specific neoglycoprotein, the so-called natural trisaccharide-octyl-bovine serum albumin (NT-O-BSA). Special features of the synthetic strategy were the use of silver trifluoromethanesulfonate (triflate) to promote glycosylation, resulting in the rhamnobiose in high yield and absolute stereospecificity. The terminal 3,6-di-O-methyl-d-glucopyranosyl group was introduced after O-deallylation of the rhamnobiose. Removal of protecting groups yielded the trisaccharide hapten suitable for coupling to carrier protein. Poly(acrylamide)-gel electrophoresis of the neoglycoprotein demonstrated its purity, and subsequent immunoblotting with a monoclonal antibody directed to the terminal 3,6-di-O-methyl-β-d-glucopyranosyl epitope of the native glycolipid demonstrated its antigenicity. Comparative serological testing in enzyme-linked immunosorbent assays of NT-O-BSA, the corresponding disaccharide-containing products, and another trisaccharide-containing neoglycoprotein, O-(3,6-di-O-methyl-β-d-glucopyranosyl)-(1→4)-O-(2,3-di-O-methyl-α-l-rhamnopyranosyl)-(1→2)-(3-O- methyl-α-l-rhamnopyranosyl)-(1→4′)-oxy-(3-phenylpropanoyl)-BSA (NT-P-BSA) [Fujiwara et al., Agric. Biol. Chem., 51 (1987) 2539-2547] against sera from leprosy patients and control populations showed concordance; the presence of the innermost sugar did not contribute significantly to sensitivity or specificity. The di- and tri-saccharide-containing neoantigens, on account of ready availability and solubility, provide greater flexibility than the native glycolipid for the serodiagnosis of leprosy.",
author = "Delphi Chatterjee and Cho, {Sang Nae} and Carol Stewart and Douglas, {James T.} and Tsuyoshi Fujiwara and Brennan, {Patrick J.}",
year = "1988",
month = "12",
day = "1",
doi = "10.1016/0008-6215(88)84078-3",
language = "English",
volume = "183",
pages = "241--260",
journal = "Carbohydrate Research",
issn = "0008-6215",
publisher = "Elsevier BV",
number = "2",

}

Synthesis and immunoreactivity of neoglycoproteins containing the trisaccharide unit of phenolic glycolipid I of Mycobacterium leprae. / Chatterjee, Delphi; Cho, Sang Nae; Stewart, Carol; Douglas, James T.; Fujiwara, Tsuyoshi; Brennan, Patrick J.

In: Carbohydrate Research, Vol. 183, No. 2, 01.12.1988, p. 241-260.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Synthesis and immunoreactivity of neoglycoproteins containing the trisaccharide unit of phenolic glycolipid I of Mycobacterium leprae

AU - Chatterjee, Delphi

AU - Cho, Sang Nae

AU - Stewart, Carol

AU - Douglas, James T.

AU - Fujiwara, Tsuyoshi

AU - Brennan, Patrick J.

PY - 1988/12/1

Y1 - 1988/12/1

N2 - The trisaccharide segment, O-(3,6-di-O-methyl-β-d-glucopyranosyl)-(1→4)-O-(2,3-di-O-methyl-α-l-rhamnopyranosyl)-(1→2)- 3-O-methyl-l-rhamnopyranose, of the Mycobacterium leprae-specific phenolic glycolipid I has been synthesized as its 8-(methoxycarbonyl)octyl glycoside and coupled to a carrier protein, to produce a leprosy-specific neoglycoprotein, the so-called natural trisaccharide-octyl-bovine serum albumin (NT-O-BSA). Special features of the synthetic strategy were the use of silver trifluoromethanesulfonate (triflate) to promote glycosylation, resulting in the rhamnobiose in high yield and absolute stereospecificity. The terminal 3,6-di-O-methyl-d-glucopyranosyl group was introduced after O-deallylation of the rhamnobiose. Removal of protecting groups yielded the trisaccharide hapten suitable for coupling to carrier protein. Poly(acrylamide)-gel electrophoresis of the neoglycoprotein demonstrated its purity, and subsequent immunoblotting with a monoclonal antibody directed to the terminal 3,6-di-O-methyl-β-d-glucopyranosyl epitope of the native glycolipid demonstrated its antigenicity. Comparative serological testing in enzyme-linked immunosorbent assays of NT-O-BSA, the corresponding disaccharide-containing products, and another trisaccharide-containing neoglycoprotein, O-(3,6-di-O-methyl-β-d-glucopyranosyl)-(1→4)-O-(2,3-di-O-methyl-α-l-rhamnopyranosyl)-(1→2)-(3-O- methyl-α-l-rhamnopyranosyl)-(1→4′)-oxy-(3-phenylpropanoyl)-BSA (NT-P-BSA) [Fujiwara et al., Agric. Biol. Chem., 51 (1987) 2539-2547] against sera from leprosy patients and control populations showed concordance; the presence of the innermost sugar did not contribute significantly to sensitivity or specificity. The di- and tri-saccharide-containing neoantigens, on account of ready availability and solubility, provide greater flexibility than the native glycolipid for the serodiagnosis of leprosy.

AB - The trisaccharide segment, O-(3,6-di-O-methyl-β-d-glucopyranosyl)-(1→4)-O-(2,3-di-O-methyl-α-l-rhamnopyranosyl)-(1→2)- 3-O-methyl-l-rhamnopyranose, of the Mycobacterium leprae-specific phenolic glycolipid I has been synthesized as its 8-(methoxycarbonyl)octyl glycoside and coupled to a carrier protein, to produce a leprosy-specific neoglycoprotein, the so-called natural trisaccharide-octyl-bovine serum albumin (NT-O-BSA). Special features of the synthetic strategy were the use of silver trifluoromethanesulfonate (triflate) to promote glycosylation, resulting in the rhamnobiose in high yield and absolute stereospecificity. The terminal 3,6-di-O-methyl-d-glucopyranosyl group was introduced after O-deallylation of the rhamnobiose. Removal of protecting groups yielded the trisaccharide hapten suitable for coupling to carrier protein. Poly(acrylamide)-gel electrophoresis of the neoglycoprotein demonstrated its purity, and subsequent immunoblotting with a monoclonal antibody directed to the terminal 3,6-di-O-methyl-β-d-glucopyranosyl epitope of the native glycolipid demonstrated its antigenicity. Comparative serological testing in enzyme-linked immunosorbent assays of NT-O-BSA, the corresponding disaccharide-containing products, and another trisaccharide-containing neoglycoprotein, O-(3,6-di-O-methyl-β-d-glucopyranosyl)-(1→4)-O-(2,3-di-O-methyl-α-l-rhamnopyranosyl)-(1→2)-(3-O- methyl-α-l-rhamnopyranosyl)-(1→4′)-oxy-(3-phenylpropanoyl)-BSA (NT-P-BSA) [Fujiwara et al., Agric. Biol. Chem., 51 (1987) 2539-2547] against sera from leprosy patients and control populations showed concordance; the presence of the innermost sugar did not contribute significantly to sensitivity or specificity. The di- and tri-saccharide-containing neoantigens, on account of ready availability and solubility, provide greater flexibility than the native glycolipid for the serodiagnosis of leprosy.

UR - http://www.scopus.com/inward/record.url?scp=0024196686&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024196686&partnerID=8YFLogxK

U2 - 10.1016/0008-6215(88)84078-3

DO - 10.1016/0008-6215(88)84078-3

M3 - Article

C2 - 3063383

AN - SCOPUS:0024196686

VL - 183

SP - 241

EP - 260

JO - Carbohydrate Research

JF - Carbohydrate Research

SN - 0008-6215

IS - 2

ER -