A modified synthesis and serological evaluation of neoglycoproteins containing the natural disaccharide of PGL-I from Mycobacterium leprae.
In order to generate substantial amounts of neoglycoconjugate needed for commercialization of diagnostic kits and high-throughput detection of leprosy, we developed a facile and high-yield synthesis of the corresponding disaccharide. Herein, the non-reducing disaccharide segment of phenolic glycolipid I from Mycobacterium leprae, O-(3,6-di-O-methyl-beta-D-glucopyranosyl)-(1-->4)-O-2,3-di-O-methyl-alpha-L-rhamnopyranose was synthesized by an improved procedure. The disaccharide was efficiently conjugated to bovine/human serum albumin, via acyl-azide intermediate, to form natural disaccharide-BSA/HSA neoglycoproteins that showed a high activity in serodiagnosis of leprosy. The disaccharide incorporated into the proteins was accurately measured by MALDI-TOF mass spectrometry. The serological activities of the neoglycoproteins against pooled human lepromatous leprosy sera were measured by ELISA and they were detectable at picogram amounts.
Zhang
Jian
J
Mycobacteria Research Laboratories, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523-1682, USA. jian.zhang@adatech.com
Chatterjee
Delphi
D
Brennan
Patrick J
PJ
Spencer
John S
JS
Liav
Avraham
A
D000942:Antigens, Bacterial / Q000737:chemistry*; D002240:Carbohydrate Sequence; D004187:Disaccharides / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay; D006017:Glycolipids / Q000737:chemistry*; D006023:Glycoproteins / Q000097:blood / Q000138:chemical synthesis* / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000942:Antigens, Bacterial; D004187:Disaccharides; D006017:Glycolipids; D006023:Glycoproteins; C045169:phenolic glycolipid I, Mycobacterium leprae
Science. 1998 Dec 11;282(5396):2076-9
9851927
Infect Immun. 1984 Jan;43(1):245-52
6360898
J Biol Chem. 1982 Dec 25;257(24):15072-8
6184369
Carbohydr Res. 1987 Jun 1;163(1):41-52
3300972
Cell. 2000 Oct 27;103(3):511-24
11081637
Carbohydr Res. 1988 Dec 1;183(2):241-60
3063383
Carbohydr Res. 1986 Nov 15;156:39-56
3815408
Carbohydr Res. 1986 May 1;148(2):287-98
3518927
Carbohydr Res. 1986 Jul 1;149(2):C13-6
3530456