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
- 批准号:6162257
- 负责人:
- 金额:--
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:
- 资助国家:美国
- 起止时间:至
- 项目状态:未结题
- 来源:
- 关键词:DNA footprinting HIV envelope protein gp120 HIV infections antibody receptor biomarker blood chemistry cancer risk epitope mapping hepatitis human immunodeficiency virus mass spectrometry monoclonal antibody neoplasm /cancer immunodiagnosis neutralizing antibody protein sequence protein structure recombinant virus virus protein
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
期刊论文数量(0)
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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:
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: