       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).

