[Analysis of antigenic determinant profiles of the Ebola virus VP35 protein N-terminal region using its short recombinant fragments].
cDNA of fragments of gene VP35 of the Ebola virus (EV) were expressed in vector pQE30 for the purpose of isolation of recombinant fragments of protein VP35. Five short affinity-purified fragments of the EV VP35 protein were analyzed, by using the methods of IEA and immunoblotting, with polyclonal antiviral sera (PAS) against EV and with hybrid monoclonal antibodies (Mabs) IC6 and 6F7 specific to EV VP35 protein. All fragments of protein VP35 with an intact N-terminal region and removed C-terminal region were found to interact effectively with PAS and with Mabs IC6 and 6F7. Rec86N, the smallest of the above fragments, comprised the initial 86 amino acid residues of the VP35 N-terminal region. A removal of 36 amino acid residues from the N-terminal region of Rec310N, the largest recombinant fragment, resulted in a loss of interaction with Mabs IC6 and 6F7, while the interaction with polyclonal antibodies remained intact. The obtained results show that the initial 86 amino acid residues of the N-terminal region of EV VP35 are of the key importance in forming the antigenic structure of VP35 and that they contain multiple B-cell epitopes. Finally, the initial 36 amino acids of VP35 predetermine the shaping-up of two antigenic determinants for Mabs IC6 and 6F7.
Rudzevich
T N
TN
Ternovoĭ
V A
VA
Kazachinskaia
E I
EI
Razumov
I A
IA
Chepurnov
A A
AA
Loktev
V B
VB
Netesov
S V
SV
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral / Q000032:analysis* / Q000276:immunology; D018076:DNA, Complementary / Q000096:biosynthesis; D029043:Ebolavirus / Q000378:metabolism*; D018604:Epitope Mapping; D000939:Epitopes / Q000032:analysis* / Q000235:genetics; D007106:Immune Sera; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D010446:Peptide Fragments / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis; D014764:Viral Proteins / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D054334:Viral Regulatory and Accessory Proteins
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D018076:DNA, Complementary; D000939:Epitopes; D007106:Immune Sera; D010446:Peptide Fragments; D011994:Recombinant Proteins; C080980:VP35 protein, filovirus; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins