Genetic Systems Corporation

Bio-Rad Laboratories, Inc.

Genetec Systems Corporation

Nobuo Monji

Carol-Ann Cole

Allan Sachs Hoffman

John H Priest

Patrick F Coleman

Raymond L Houghton

Robert C Nowinski

James J Blake

Peter Chong-Dug Su

John P Montana

Alice Kamp Goshorn

Novel polypeptides are provided having substantially the same sequence as immunologically significant fragments of AIDS-related viruses. The polypeptides can be used as reagents in the determination of exposure of a human host to the virus. Of particular interest is the use of polypeptides in screening blood products.

Synthetic antigen for the detection of antibodies immunoreactive with HIV virus

Peptides immunoreactive with antibodies to native proteins, and which have at least two cysteine residues that contribute to mimicking an epitope of the protein, are prepared with the cysteine thiol groups protected. When deprotected, the peptides have enhanced immunoreactivity. The peptides are particularly useful for detecting antibodies or antigens associated with retroviruses, including the clinically important lymphotropic retroviruses HIV-1, HIV-2, HTLV-I, and HTLV-II.

Cysteine thiol-protected peptides for use in immunoassays

Methods for determining the presence and/or concentration of an analyte in a biological fluid sample are disclosed. The methods generally include admixing in solution certain polymer/reactant and reporter/reactant conjugates along with the biological fluid sample suspected of containing the analyte, thereby forming ternary complexes. The separation of the complexes from the reaction mixture is achieved through the affinity of certain selected polymer compositions for various solid phases. Upon separation, the amount of reporter activity in the solution may be measured, and therefrom the presence and/or concentration of the analyte determined. Multiple analyses on a biological fluid sample suspected of containing one or more analytes may also be performed, using either a variety of different reporters or selected polymers having varied affinity for the solid phase.

Membrane affinity concentration immunoassay

Rapid assays for analytes of interest in a fluid sample utilize a first conjugate of a labelled reactant that specifically binds to the analyte, and a second conjugate that binds to the analyte coupled to a polymer that has an affinity for a selected solid phase. The reaction components are incubated briefly, then contacted with the selected solid phase and the labelled components determined. Optional wash steps provide for enhanced sensitivity and specificity. When the analyte of interest is an antibody to HIV, the first reactant may be a synthetic, recombinant or native HIV antigen, and the second reactant may be protein A or an anti-immunoglobulin.

Rapid membrane affinity concentration assays

Methods for delivering substances into, removing substances from, or reacting substances with a selected environment utilizing polymer gels or coatings characterized by a critical solution temperature (CST) are disclosed. The CST as well as the pore structure, pore size, pore distribution, and absorbing capacity of the gel may be selectively controlled. The substances may be physically or chemically immobilized within the polymer gels. In addition, a method for altering the surface wettability of CST polymers is also disclosed.

Methods for selectively reacting ligands immobilized within a

An immunoassay in which a thermally induced phase separation is used to effect the separation of specifically bound reactants from free reactants is disclosed. A first reactant is conjugated to a temperature-sensitive polymer to form a polymer/reactant conjugate, and a second reactant is conjugated to a reporter to form a reporter/reactant conjugate. The polymer/reactant, reporter/reactant, and biological fluid samples suspected of containing the analyte are admixed in solution at a temperature other than that at which the polymer will precipitate. Specific binding is allowed to occur, thereby forming a ternary complex. The salt concentration of the adjusted solution is then adjusted to a concentration sufficient to cause the complex to precipitate from the solution, the amount of reporter activity in the precipitated complex or in the solution measured and the presence and/or concentration of the analyte therefrom determined. Alternatively, the first reactant may be conjugated to a monomer and subsequently copolymerized with additional monomers to yield a temperature-sensitive copolymer. Multiple analyses may also be performed on a single sample by choosing a variety of polymers, each polymer having a different specific binding partner conjugated thereto and a different critical solution temperature. By altering the temperature and/or the salt concentration of the solution incrementally, the reporter associated with each of the complexes precipitated with each temperature or concentration increment may be measured, and the presence and/or concentration of each of the analytes determined.

Thermally induced phase separation immunoassay

Conjugate monomers, polymers, and methods for the de novo synthesis of the polymers are provided. Conjugate organic monomers contain binding-pair members which upon polymerization become integrally associated with the resultant polymer. Specifically, antigens, antibodies, receptors, and ligands may be bound to organic monomers either directly by chemical reaction or indirectly by chemical spacer arms, and these conjugates may be polymerized or copolymerized with nonderivatized monomers to form polymers containing variable amounts of the binding-pair members. Such conjugate monomers and polymers find a wide variety of uses in binding to their binding-pair-member cognate which include selective removal of complementary binding-pair members from solution as well as in immunoassay procedures and in immunization regimes.

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Seattle, WA, United States of America

Nobuo Monji