Classifications

• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
  • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
  • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
  • G01N33/56983—Viruses
  • G01N33/56988—HIV or HTLV
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
  • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
  • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
  • G01N33/56983—Viruses

Definitions

• This invention relates to a process to coat viral antigens in the presence of nonionic detergent and acidic pH, onto a solid phase, to enhance the activity of bound viral antigens.
• Antibodies are immobilized on solid phases, such as microtiter plates and latex beads, for use in research and diagnostic assays. Detection of monoclonal antibodies to membrane-associated proteins, however, presents a problem, since nonionic detergents used to
• This invention relates to a method to increase the concentration of viral antigen on a solid phase by coating the solid phase with lu viral antigen in the presence of nonionic detergents, such as Triton X-l ⁇ U or NP4U, in acidic pH.
• nonionic detergents such as Triton X-l ⁇ U or NP4U
• Antigen was diluted into a coating buffer and added to lb particles.
• the coating buffer was preferably comprised of 0.1 M acetate, pH b.O with 5* NP40, however, other nonionic detergents, such as Triton X-100, can be used. It was observed, however, that one nonionic detergent, Tween, did not result in a sufficiently high concentration of antigen to conduct an immunoassay.
• 2U tion of detergent can range from .2% to 10 , with the optimum concentration from between about 1 and 10%. It was observed that pH can range from 4.U to 6.U, with the optimum pH at 5.0.
• Buffer A was comprised of b 0.1 M acetate, at a pH 5.0.
• Buffer B was comprised of 0.1 M acetate, 0.1 M NaCl, at a pH 5.0.
• Buffer C was comprised of 0.1 M acetate, 0.1 M NaCl, 0.2% NP40, at a pH 5.0.
• HIV negative serum was designated "617", positive was designated “618", and high positive serum was designated “R-J 1825”.
• SDB sample dilution buffer
• the dialyzed preparation was coated at 1/2 dilution with coating buffers A, B, and C on to 1 ml of 2.5% carboxylated paramagnetic particles (3.5 urn). Particles were incubated overnight at 2 Q 8 lb C. The particles were pelleted, the supernatent was discarded, and the particles were resuspended in 1% normal goat serum (NGS) in 0.1 M acetate, at a pH 5.0.
• NGS normal goat serum
• the coated particles were incubated for two hours at room temperature, then washed three times with 1 L isotonic buffered U saline (IBS).
• IBS isotonic buffered U saline
• the coating buffer "C” comprised of 0.1 M acetate, 0.1 M NAC1 , 30 and 0.2A NP40, and resulted in a greater concentration of viral antigens on the solid phase.
• AIDS antigens coated in the presence of nonionic detergents and acidic pH resulted in viral coated particles witn enhanced activity over particles coated using other buffers.

Landscapes

• Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Immunology (AREA)
• Engineering & Computer Science (AREA)
• Molecular Biology (AREA)
• Hematology (AREA)
• Virology (AREA)
• Biomedical Technology (AREA)
• Chemical & Material Sciences (AREA)
• Urology & Nephrology (AREA)
• Food Science & Technology (AREA)
• Biochemistry (AREA)
• Pathology (AREA)
• Cell Biology (AREA)
• Biotechnology (AREA)
• Tropical Medicine & Parasitology (AREA)
• Medicinal Chemistry (AREA)
• Physics & Mathematics (AREA)
• Analytical Chemistry (AREA)
• Microbiology (AREA)
• General Health & Medical Sciences (AREA)
• General Physics & Mathematics (AREA)
• AIDS & HIV (AREA)
• Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
• Detergent Compositions (AREA)
• Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
• Peptides Or Proteins (AREA)

Applications Claiming Priority (2)

Publications (2)

Family

ID=23761588

Family Applications (1)

Country Status (5)

Families Citing this family (1)

Citations (3)

Family Cites Families (1)

• 1990
  • 1990-11-28 CA CA 2045682 patent/CA2045682A1/en not_active Abandoned
  • 1990-11-28 EP EP19910902536 patent/EP0455807A4/en not_active Withdrawn
  • 1990-11-28 JP JP50267491A patent/JPH04507003A/ja active Pending
  • 1990-11-28 AU AU71604/91A patent/AU7160491A/en not_active Abandoned
  • 1990-11-28 WO PCT/US1990/006922 patent/WO1991008311A1/en not_active Application Discontinuation

Patent Citations (3)

Non-Patent Citations (2)

Also Published As

Similar Documents

Legal Events