Characterization of conformational epitope of alginate-derived polymannuronates by surface plasmon resonance.
In the present study, we developed a mAb to alginate-derived polymannuronates (ADPM) and examined the antigenic epitopes using surface plasmon resonance (SPR) in conjunction with a large panel of oligomannuronate probes. We found that tetrasaccharide is the minimum-binding unit, and that increases in chain length from the tetrasaccharide to the heptsaccharide further enhance monovalent binding. A sharp increase in affinity was observed when increasing from the octasaccharide to the cosasaccharide, which is due to a further enhancement of the individual antigenic epitope combined with multivalency. Kinetic binding studies further suggested that the conformational epitope is discontinuous and infrequent and that a C6-carboxyl group is important in maintaining the conformational epitope. Moreover, CD analysis revealed there were conformational structures in epitopes. The data support our hypothesis that the conformational epitope for the mAb may be an extended helical segment of ADPM. ADPM exists mainly in linear form, but it can infrequently and spontaneously form extended helices. Although helices are not prevalent in ADPM, the immune system preferentially selects these conformational epitopes because they are unique. Together, our results indicate that the antigenic epitopes in beta-d-mannuronates are conformational and require C6-carboxyl groups.
Gan
Li
L
Department of Pharmacology, Marine Drug and Food Institute, Ocean University of China, 5 Yushan Road, Qingdao 266003, China.
Xin
Xian L
XL
Li
Jing
J
Liu
Hai Y
HY
Zhang
Zhen Q
ZQ
Qian
Jin
J
Ding
Jian
J
Geng
Mei Y
MY
D000464:Alginates / Q000737:chemistry*; D000818:Animals; D000911:Antibodies, Monoclonal; D002236:Carbohydrate Conformation; D002942:Circular Dichroism; D000939:Epitopes / Q000737:chemistry*; D008351:Mannans / Q000737:chemistry*; D051379:Mice; D009844:Oligosaccharides / Q000737:chemistry; D020349:Surface Plasmon Resonance
D000464:Alginates; D000911:Antibodies, Monoclonal; D000939:Epitopes; D008351:Mannans; D009844:Oligosaccharides