Isolation of recombinant partial gag gene product p18 (HIV-1Bru) from Escherichia coli

Kolbe, Hanno V. J.; Jaeger, Francine; Lepage, Pierre; Roitsch, C.; Lacaud, Georges; Kiény, Marie-Paule; Sabatié, J.; Brown, Stephen; Lecocq, Jean‐Pierre; Girard, Marc

doi:10.1016/s0021-9673(01)93860-9

Cited by 5 publications

(2 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Soluble recombinant proteins p27 nef and p18 gag were purified from Escherichia coli (25,36). The p27 nef -V3 conjugate was prepared by chemical N-terminal tyrosine linkage, as described before (21).…”

Section: Methodsmentioning

confidence: 99%

Vaccine-induced protection of chimpanzees against infection by a heterologous human immunodeficiency virus type 1

Girard

Meignier

Barré‐Sinoussi

et al. 1995

J Virol

View full text Add to dashboard Cite

The extraordinary genetic diversity of human immunodeficiency virus type 1 (HIV-1) is a major problem to overcome in the development of an effective vaccine. In the most reliable animal model of HIV-1 infection, chimpanzees were immunized with various combinations of HIV-1 antigens, which were derived primarily from the surface glycoprotein, gp160, of HIV-1 strains LAI and MN. The immunogens also included a live recombinant canarypox virus expressing a gp160-MN protein. In one experiment, two chimpanzees were immunized multiple times; one animal received antigens derived only from HIV-1 LAI , and the second animal received antigens from both HIV-1 LAI and HIV-1 MN . In another experiment, four chimpanzees were immunized in parallel a total of five times over 18 months; two animals received purified gp160 and V3-MN peptides, whereas the other two animals received the recombinant canarypox virus and gp160. At 3 months after the final booster, all immunized and naive control chimpanzees were challenged by intravenous inoculation of HIV-1 SF2 ; therefore, the study represented an intrasubtype B heterologous virus challenge. Virologic and serologic follow-up showed that the controls and the two chimpanzees immunized with the live recombinant canarypox virus became infected, whereas the other animals that were immunized with gp160 and V3-MN peptides were protected from infection. Evaluation of both cellular and humoral HIV-specific immune responses at the time of infectious HIV-1 challenge identified the following as possible correlates of protection: antibody titers to the V3 loop of MN and neutralizing antibody titers to HIV-1 MN or HIV-1 LAI , but not to HIV-1 SF2 . The results of this study indicate that vaccine-mediated protection against intravenous infection with heterologous HIV-1 strains of the same subtype is possible with some immunogens.

show abstract

Section: Methodsmentioning

confidence: 99%

Vaccine-induced protection of chimpanzees against infection by a heterologous human immunodeficiency virus type 1

Girard

Meignier

Barré‐Sinoussi

et al. 1995

J Virol

View full text Add to dashboard Cite

show abstract

“…Recombinant gpl60env was purified from the culture medium of BHK21 cells infected with VV-1163, a recombinant vaccinia virus expressing the gpl60 env gene modified by site-directed mutagenesis to destroy the gp120/41 cleavage site and to remove the anchor domain of gp4l (12,13). Where indicated, the antigen was mixed with recombinant pl8gag, p27nef, and p23vif antigens that were purified from Escherichia coli pTG2153, pTG1166, and pTG1149, respectively, as described (14,15 (21). CD4+-enriched lymphocytes were obtained from chimpanzee PBMC by separation with magnetic beads to which were attached monoclonal antibodies specific for the CD8 antigen (Dynabeads, Robbins Scientific, Mountain View, CA).…”

Section: Methodsmentioning

confidence: 99%

Immunization of chimpanzees confers protection against challenge with human immunodeficiency virus.

Girard

Kiény

Pinter

et al. 1991

Proc. Natl. Acad. Sci. U.S.A.

310

148

View full text Add to dashboard Cite

Sustained high titers of neutralizing antibodies were elicited in three chimpanzees after sequential injections of different human immunodeficiency virus 1 (HIV-1) antigen preparations derived from the HIV-1 BRU strain that included whole inactivated virus or purified recombinant proteins and then synthetic peptides identical to the major HIV-1 neutralizing epitope V3. The animals were chaflenged i.v. with 40 chimpanzee infectious doses (equivalent to 100 tissue culture 50% infectious doses) of a stock of HIV-1 IIIB isolate. After 6 mo of follow-up, all three animals appeared uninfected by serologic and virologic criteria, including polymerase chain reaction analysis and failure to isolate virus from peripheral blood lymphocytes, bone marrow, and lymph node tissue. Of two chimpanzees monitored for 1 yr, virus was isolated initially from one animal at 32 weeks, but the second chimpanzee was virus negative by all assays through 12 mo; the third animal has remained virus negative through 9 mo of follow-up. These results indicate that it is possible to elicit protection against, or significantly delay infection of, HIV-1 by immunization, thus laying the foundation for development of an HIV-1 vaccine.

show abstract

Application of electrospray mass spectrometry to the characterization of recombinant proteins up to 44 kDa

Dorsselaer

Bitsch

Green³

et al. 1990

Biol. Mass Spectrom.

Self Cite

View full text Add to dashboard Cite

Mass measurement by electrospray mass spectrometry (ESMS) is used as a rapid preliminary verification of the identity of various recombinant proteins ranging from 7 to 44 kDa with an accuracy of 0.01-0.03%. ESMS not only improves the speed but also the reliability of the protein structure determination when used in conjunction with other methods of protein analysis. Modifications of these large molecules, for example the loss of C-terminal amino acids, N-terminal acetylation, 2-mercaptoethanol addition to a cysteine, and trace formation of a covalent dimer (3%), are easily detected individually or in mixtures by mass measurement using ESMS; feats which would be very difficult to achieve using classical biochemical methods. As little as 1% of several structurally related protein contaminants have been identified in a 15 kDa recombinant protein preparation.

show abstract

Isolation of recombinant partial gag gene product p18 (HIV-1Bru) from Escherichia coli

Cited by 5 publications

References 11 publications

Vaccine-induced protection of chimpanzees against infection by a heterologous human immunodeficiency virus type 1

Vaccine-induced protection of chimpanzees against infection by a heterologous human immunodeficiency virus type 1

Immunization of chimpanzees confers protection against challenge with human immunodeficiency virus.

Application of electrospray mass spectrometry to the characterization of recombinant proteins up to 44 kDa

Contact Info

Product

Resources

About