ANTISERA ASSAYS FOR MLV RELATED VIRUSES IN HUMANS AND OTHER MAMMALS
The disclosure provide cell lines and methods for the production of vectors and viral particles useful in diagnostics and therapeutics.
1 . A method for detecting anti-viral antibodies for amphotropic MLV or MLV like virus in a biological sample compromising
a) incubating a biological sample with a capture reagent immobilized on a solid support to bind multiple anti-viral antibodies to amphotropic MLV to the capture reagent, wherein the capture reagent comprises a purified viral preparation;
b) detecting bound anti-viral antibodies bound to immobilized capture reagent by contacting the bound anti-viral antibodies with a labeled detecting secondary agent.
2 . The method of claim 1 , wherein the biological sample is isolated from a mammal.
3 . The method of claim 2 , wherein the mammal is an autoimmune, virally infected, immune suppressed or cancer patient.
4 . The method of claim 1 , wherein the biological sample is isolated from a human.
5 . The method of claim 4 , wherein the human is an autoimmune, virally infected immune suppression or cancer patient.
6 . The method of claim 5 , further comprising measuring the amount of anti-viral antibody, wherein the amount of antibody is titred using a dilution series; wherein the amount of antibody determined is IgG or IgM by use of specific detecting secondary reagents.
7 . The method of claim 1 , wherein the biological sample is plasma, serum, urine, saliva, buccal swab, nasal swab, fine needle aspirate, semen, vaginal secretion or milk.
8 . The method of claim 1 , wherein the capture reagent is immobilized at about 1.5 μg/mL.
9 . The method of claim 1 , wherein the solid support is a microtiter plate, a glass slide, a nylon slide, or a PVDF membrane.
10 . The method of claim 1 , wherein the solid support is a latex bead or a microsphere.
11 . The method of claim 1 , wherein the solid support is a microfluidic chip or a semiconductor chip.
12 . The method of claim 1 , wherein the secondary agent is a secondary antibody that is detectably labeled.
13 . The method of claim 1 , further comprising comparing the detected bound anti-viral antibodies to a control comprising human serum or plasma seropositive for MLV.
14 . The method of claim 13 , further comprising identifying the control with the monoclonal 83A25 mAb.
15 . The method of claim 1 , wherein the assay is used for diagnostic purposes.
16 . An immunoassay kit for detecting anti-viral antibodies comprising:
a) an antibody that specifically binds to amphotropic envelop as a control;
b) a capture reagent comprising a purified viral particle.
17 . A kit of claim 16 , wherein the solid support is a microtiter plate.
18 . A kit of claim 16 , wherein the solid support is a microsphere.
19 . A kit of claim 16 , wherein the capture reagent is an MLV-like viral particle.
20 . A kit of claim 16 , wherein the capture reagent is a purified protein of MLV origin.
21 . A kit of claim 16 , wherein the capture reagent is a purified recombinant protein of MLV origin or MLV-like sequence.
22 . A kit of claim 16 , wherein the detectable antibody is a monoclonal antibody.
23 . The kit of claim 22 , wherein the monoclonal antibody is a murine monoclonal antibody.
24 . The kit of claim 16 , wherein the antibody is a polyclonal antibody.
25 . The kit of claim 24 , wherein the polyclonal antibody is a rabbit or goat, sheep or chicken polyclonal antibody.
26 . The kit of claim 16 , wherein the kit is used in a clinical or diagnostic setting.
27 . The kit of claim 16 , wherein the kit is used to monitor the immune responses of MLV-infected or MLV-like infected individuals.
28 . The method of claim 1 , wherein the purified viral particle is produced by a method comprising:
transforming a 293 cell line with a plasmid encoding a retroviral MLV or MVL-like virus;
culturing the 293 cell to produce viral particles;
isolating the viral particles;
infecting an HT1080 cell line with the viral particles thereby producing the viral particle producing cell line, producing the viral particles and purifying the viral particles.
29 . The kit of claim 16 , wherein the purified viral particle is produced by a method comprising:
transforming a 293 cell line with a plasmid encoding a retroviral MLV or MVL-like virus;
culturing the 293 cell to produce viral particles;
isolating the viral particles;
infecting an HT1080 cell line with the viral particles thereby producing the viral particle producing cell line, producing the viral particles and purifying the viral particles.