Monoclonal antibodies to HIV-1 and methods of using same
Inventors
Kopp, Jeffrey B. • Phillips, Terence M. • Schubert, Ulrich • Yewdell, John
Assignees
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Publication Number
US-7993647-B2
Publication Date
2011-08-09
Expiration Date
2025-06-22
Abstract
The present invention provides monoclonal antibodies to HIV-1 Vpr and hybridoma cell lines that produce the monoclonal antibodies to HIV-1 Vpr. Methods for use of such antibodies in the detection of HIV-1 infection are also provided.
Core Innovation
The invention provides monoclonal antibodies to the HIV-1 viral protein R (Vpr) and hybridoma cell lines that produce these monoclonal antibodies. It also includes methods for using such antibodies to detect HIV-1 infection by identifying the presence of Vpr in biological fluids. The antibodies described include monoclonal, chimeric, and humanized forms and can be conjugated to detectable labels for use in immunoassays.
The problem addressed is the lack of effective means to detect or diagnose complications of HIV-1 infection that may arise from the action of HIV-1 regulatory or accessory proteins like Vpr on infected or bystander cells. Existing immunoassays detect HIV-1 antibodies or monitor disease progression via CD4 counts and viral RNA levels, but no clinical assay measures levels of HIV-1 regulatory proteins in biological fluids, which may correlate with HIV-1 complications such as neuropathy, dementia, metabolic syndrome, or nephropathy.
Claims Coverage
The patent includes several independent claims covering antibodies, hybridoma cell lines, and methods for detecting HIV-1 Vpr. The claims encompass both compositions and diagnostic methods involving specific antibodies and detection techniques.
Antibody competitively inhibiting binding of a second antibody produced by hybridoma 9F12
A first antibody to HIV-1 viral protein R that competitively inhibits binding of a second antibody produced by hybridoma cell line 9F12. The first antibody includes the six complementarity determining regions (CDRs) from the second antibody and can be a monoclonal, chimeric, humanized antibody, or a functional fragment comprising the six CDRs.
Isolated hybridoma cell line 9F12 producing anti-Vpr antibody
An isolated hybridoma cell line 9F12 deposited as PTA-5560, which produces the monoclonal antibody to HIV-1 Vpr.
Method for detection of HIV-1 viral protein R in a sample using a first antibody
A method for detection of HIV-1 viral protein R involving (i) contacting a sample with a first antibody comprising the six CDRs from the second antibody produced by hybridoma 9F12 that competitively inhibits binding of the second antibody, and (ii) detecting the antibody-Vpr complex formed in the sample.
Quantitation and comparison of HIV-1 Vpr to a standardized control for disease progression and complications
A method including quantitating the amount of the antibody-Vpr complex to determine the level of HIV-1 Vpr in a biological fluid sample, and comparing this amount to a standardized control to determine HIV-1 disease progression or presence of HIV-1 associated complications such as neuropathy, dementia, metabolic syndrome, or nephropathy.
Detectable labeling and immuno-capillary electrophoresis detection
The first antibody or monoclonal antibody may be conjugated to a detectable label, and detection of the antibody-Vpr complex in the method can be performed using capillary electrophoresis or immunoaffinity capillary electrophoresis.
The claims cover monoclonal and derivative antibodies specifically targeting HIV-1 viral protein R produced by a known hybridoma, isolated hybridoma cell lines themselves, and methods for sensitive detection and quantitation of HIV-1 Vpr in biological samples. The methods include labeling and advanced detection techniques and allow for assessing disease progression and specific HIV-1 complications.
Stated Advantages
Provides reliable, sensitive systems for detection of HIV-1 infection through detection of Vpr, which functions throughout the viral life cycle.
Enables measurement of HIV-1 regulatory proteins like Vpr in biological fluids, which may correlate with HIV-1 complications, thus aiding diagnosis and management of such complications.
Allows early diagnosis of HIV-1 complications, potentially improving patient quality of life and disease management choices.
Supports development and monitoring of new therapies targeting Vpr or its effects on host cells.
Documented Applications
Detection of HIV-1 infection by identifying HIV-1 viral protein R (Vpr) in biological fluids such as plasma and serum.
Diagnosis and monitoring of HIV-1 disease progression by quantitating Vpr levels and comparing to standardized controls.
Detection and diagnosis of complications associated with HIV-1 infection including neuropathy, dementia, metabolic syndrome, and nephropathy through measurement of Vpr in biological fluids.
Use of immunoaffinity capillary electrophoresis assays employing immobilized monoclonal antibodies for sensitive quantitation of Vpr in patient samples.
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