A simple one-step method for the preparation of HIV-1 envelope glycoprotein immunogens based on a CD4 mimic peptide.
To counteract the problems associated with the purification of HIV envelope, we developed a new purification method exploiting the high affinity of a peptide mimicking CD4 towards the viral glycoprotein. This miniCD4 was used as a ligand in affinity chromatography and allowed the separation in one step of HIV envelope monomer from cell supernatant and the capture of pre-purified trimer. This simple and robust method of purification yielded to active and intact HIV envelopes as proved by the binding of CCR5 HIV co-receptor, CD4 and a panel of well-characterized monoclonal antibodies. The immunogenicity of miniCD4-purified HIV envelope was further assessed in rats. The analysis of the humoral response indicated that elicited antibodies were able to recognize a broad range of HIV envelopes. Finally, this method based on a chemically synthesized peptide may represent a convenient and versatile tool for protein purification compatible far scale-up in both academic and pharmaceutical researches.
Martin
Grégoire
G
CEA, iBiTecS, Service d'Ingénierie Moléculaire des Protéines, Bât 152, Gif sur Yvette, F-91191, France.
Sun
Yide
Y
Heyd
Bernadette
B
Combes
Olivier
O
Ulmer
Jeffrey B
JB
Descours
Anne
A
Barnett
Susan W
SW
Srivastava
Indresh K
IK
Martin
Loïc
L
D000818:Animals; D015704:CD4 Antigens / Q000378:metabolism*; D016466:CHO Cells; D002846:Chromatography, Affinity / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D015483:HIV Antibodies / Q000378:metabolism; D015699:HIV Envelope Protein gp120 / Q000276:immunology / Q000302:isolation & purification*; D015497:HIV-1; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D055503:Protein Multimerization; D051381:Rats; D017208:Rats, Wistar; D019713:Receptors, CCR5 / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism
D015704:CD4 Antigens; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D010455:Peptides; D019713:Receptors, CCR5; D011994:Recombinant Proteins