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M9630040.TXT
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1996-02-27
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Document 0040
DOCN M9630040
TI Efficient purification and rigorous characterisation of a recombinant
gp120 for HIV vaccine studies.
DT 9603
AU Jones DH; McBride BW; Roff MA; Farrar GH; Centre for Applied
Microbiology and Research, Salisbury,; Wiltshire, UK.
SO Vaccine. 1995 Aug;13(11):991-9. Unique Identifier : AIDSLINE
MED/96088051
AB A recombinant HIV-1 gp120 (rgp120) was expressed in a permanent Chinese
Hamster Ovary (CHO) cell line (L761h) that constitutively secretes the
product of clone p4 derived from the env gene of HIV-1 isolate GB8. The
rgp120 was isolated from cell culture supernatants by a simple, rapid,
non-denaturing and efficient purification procedure based on a novel
combination of lectin affinity and FPLC ion-exchange chromatography. The
purity of the isolated glycoprotein was rigorously confirmed by
SDS-PAGE, capillary electrophoresis, laser desorption mass spectrometry,
total amino acid analysis and N-terminal amino acid sequencing. The
retention of biological activity by the purified rgp120 was assessed by
determining the dissociation constant of rgp120 binding to sCD4. After
formulation of this highly purified and biologically active rgp120 with
conventional adjuvants, including types already used in clinical trials
of candidate gp120-based HIV vaccines, antibody responses in immunised
rabbits were analysed using panels of overlapping synthetic peptides.
The consequences of using currently available adjuvants to deliver
highly specialised and perhaps conformation-dependent molecules, like
HIV gp120, are presented and discussed in the context of HIV vaccine
development.
DE Amino Acid Sequence Animal Antigens, CD4/CHEMISTRY AIDS
Vaccines/CHEMISTRY/*IMMUNOLOGY/ISOLATION & PURIF CHO Cells
Electrophoresis, Capillary Electrophoresis, Polyacrylamide Gel
Hamsters HIV Antibodies/BIOSYNTHESIS/CHEMISTRY HIV Envelope Protein
gp120/CHEMISTRY/*IMMUNOLOGY/*ISOLATION & PURIF Molecular Sequence Data
Protein Binding Spectrum Analysis, Mass Support, Non-U.S. Gov't
Vaccines, Synthetic/CHEMISTRY/IMMUNOLOGY JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).