EPITOPE MAPPING OF HIV PROTEINS USING PROTEOLYTIC FOOT PRINTING AND MS

使用蛋白水解足印迹和 MS 对 HIV 蛋白进行表位作图

基本信息

项目摘要

Summary of Work: Mapping epitopes of the Human Immunodeficiency Virus (HIV) is important for the diagnosis of infection and for the development of vaccines and therapeutics for Acquired Immune Deficiency Syndrome (AIDS). We have been probing epitopes on the HIV proteins gp120 and p24. The initial step in the entry of HIV into the host cell is binding of the envelope glyco-protein gp120 to the cellular receptor CD4. Also gp120 elicits the major components of the protective immune response against HIV in humans and chimpanzees. gp120 and its synthetic peptides have been investigated as potential vaccine candidates. Similarly, HIV p24 elicits the first antibodies upon HIV infection. As the HIV infection progresses to AIDS, there is a simultaneous reduction in anti-p24 antibody titer. It has been proposed that a combination vaccine eliciting antibodies to both gp120 and p24 may be useful in combating HIV infection. Thus, knowledge of the antigenic determinants on p24 and gp120, especially those eliciting the formation of protective antibodies, is extremely important in the development of a vaccine.We have combined proteolytic footprinting and MALDI/MS to map epitopes on the native proteins recognized by antibodies. In this method, proteins affinity- bound to an immobilized antibody are proteolytically cleaved and the unbound fragments are removed by washing. The bound fragments containing the epitope are characterized by directly analyzing the immobilized antibody by MALDI/MS. We have identified the core epitope on recombinant HIV p26 identified by the monoclonal antibody 13-102-100 as being residues 102-112. The cyclophilin A binding region is also contained within these residues. We have also determined the antigenic determinant of the envelope glycoprotein gp120 recognized by a polyclonal antibody raised against the C-terminus of the protein. Antibodies against the C- terminus often show protective effects in vitro. We are currently mapping an epitope on gp120 recognized by a MAb obtained from sera of an HIV infected individual characterized as a slow progressor, i.e., an individual who may be producing protective antibodies. We are also mapping a discontinuous epitope on HIV p24. This latter example utilizes the proteolytic footprinting results combined with the known molecular structure to identify amino acids involved in the epitope.
工作综述:人类免疫缺陷病毒表位的定位

项目成果

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K B TOMER其他文献

K B TOMER的其他文献

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{{ truncateString('K B TOMER', 18)}}的其他基金

EPITOPE MAPPING OF HIV PROTEINS USING ASSAYS IN CONJUCTION W/MASS SEPECTROMETRY
结合质谱分析使用 HIV 蛋白表位作图
  • 批准号:
    2452862
  • 财政年份:
  • 资助金额:
    --
  • 项目类别:
COLLABORATIVE PROJECTS IN ENVIRONMENTAL HEALTH SCIENCES
环境健康科学合作项目
  • 批准号:
    5202202
  • 财政年份:
  • 资助金额:
    --
  • 项目类别:
ENVIRONMENTAL HEALTH APPLICATIONS OF MASS SPECTROMETRY
质谱法在环境健康方面的应用
  • 批准号:
    3918689
  • 财政年份:
  • 资助金额:
    --
  • 项目类别:
APPLICATION OF THERMOSPRAY LC-MS TO STRUCTURE ELUCIDATION OF BIOMOLECULES
热喷雾液质联用技术在生物分子结构解析中的应用
  • 批准号:
    3918702
  • 财政年份:
  • 资助金额:
    --
  • 项目类别:
DEVELOPMENT OF FAB/MS-MS FOR ENVIRONMENTAL HEALTH SCIENCES
环境健康科学 FAB/MS-MS 的开发
  • 批准号:
    3941541
  • 财政年份:
  • 资助金额:
    --
  • 项目类别:
STRUCTURE ELUCIDATION OF CARCINOGEN-NUCLEOSIDE ADDUCTS
致癌物-核苷加合物的结构解析
  • 批准号:
    3941543
  • 财政年份:
  • 资助金额:
    --
  • 项目类别:
DEVELOPMENT OF NANOLITER CAPILLARY LC/MS TECHNIQUES
纳升毛细管 LC/MS 技术的开发
  • 批准号:
    3841101
  • 财政年份:
  • 资助金额:
    --
  • 项目类别:
DEVELOPMENT OF NANOLITER CAPILLARY LC/MS TECHNIQUES
纳升毛细管 LC/MS 技术的开发
  • 批准号:
    3755449
  • 财政年份:
  • 资助金额:
    --
  • 项目类别:
COLLABORATIVE PROJECTS IN ENVIRONMENTAL HEALTH SCIENCES
环境健康科学合作项目
  • 批准号:
    3876935
  • 财政年份:
  • 资助金额:
    --
  • 项目类别:
MICRODIALYSIS/MASS SPECTROMETRY
微量透析/质谱分析
  • 批准号:
    3855930
  • 财政年份:
  • 资助金额:
    --
  • 项目类别:
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