Detection of Ebola virus envelope using monoclonal and polyclonal antibodies in ELISA, surface plasmon resonance and a quartz crystal microbalance immunosensor.
Ebola virus (EBOV) Zaire, Sudan, as well as Ivory Coast are virulent human EBOV species. Both polyclonal and monoclonal antibodies (MAbs) were developed against soluble EBOV envelope glycoprotein (GP) for the study of EBOV envelope diversity and development of diagnostic reagents. Three EBOV Sudan-Gulu GP peptides, from the N-terminus, mid-GP, and C-terminus regions were used to immunize rabbits for the generation of anti-EBOV polyclonal antibodies. Polyclonal antisera raised against the C-terminus peptide could detect both Sudan-Gulu as well as Zaire GPs, while anti-N and mid-region peptide polyclonal sera recognized only EBOV Sudan-Gulu GP. Of the three anti-EBOV GP mouse MAbs produced, MAb 15H10 recognized all human EBOV GP species tested (Zaire, Sudan and Ivory Coast), and as well as reacted with the Reston non-human primate EBOV GPs. In addition, MAb 15H10 bound virion-associated GP of all known EBOV species. MAb 17A3 recognized GPs of both EBOV Sudan-Gulu and Zaire, while MAb 6D11 recognized only EBOV Sudan-Gulu GP. To detect EBOV GP, these antibody reagents were used in ELISA, surface plasmon resonance and in a quartz crystal microbalance immunosensor. Thus, polyclonal and monoclonal antibodies can be used in combination to identify and differentiate both human and non-human primate EBOV GPs.
Yu
Jae-Sung
JS
Human Vaccine Institute, Duke University Medical Center, Durham, NC 27710, United States. yu000022@mc.duke.edu
Liao
Hua-Xin
HX
Gerdon
Aren E
AE
Huffman
Brian
B
Scearce
Richard M
RM
McAdams
Mille
M
Alam
S Munir
SM
Popernack
Paul M
PM
Sullivan
Nancy J
NJ
Wright
David
D
Cliffel
David E
DE
Nabel
Gary J
GJ
Haynes
Barton F
BF
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D003429:Cross Reactions; D029043:Ebolavirus / Q000145:classification / Q000276:immunology / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D020349:Surface Plasmon Resonance / Q000379:methods*; D014759:Viral Envelope Proteins / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D014759:Viral Envelope Proteins; C113189:envelope glycoprotein, Ebola virus