Fungal detection using mannan epitope
Non-invasive methods are provided herein for diagnosing samples as including a fungus, including fungal infection or contamination, with specific monoclonal antibodies capable of detecting molecules associated with fungi in the sample, such as a biological or environmental sample. These molecules can be identified using various methods, including but not limited to antibody based methods, such as an enzyme-linked immunosorbant assay (ELISA), or a lateral flow immunoassay.
1. A composition comprising a monoclonal antibody that specifically binds the α-1,6 mannose backbone of fungal mannan, wherein the antibody comprises an amino acid sequence encoded by at least a light chain nucleic acid sequence comprising SEQ ID NO: 1 of mAb 2DA6 or at least a heavy chain nucleic acid sequence comprising SEQ ID NO: 2 of mAb 2DA6.
2. The composition of claim 1 , wherein mAb 2DA6 is produced by a hybridoma having ATCC Accession No. PTA-123011.
3. A composition comprising an antibody that specifically binds the α-1,6 mannose backbone of fungal mannan, wherein the antibody is a single chain
scFv clone 11C16 comprising an amino acid sequence encoded by a light chain nucleic acid sequence comprising SEQ ID NO: 5 and a heavy chain nucleic acid sequence comprising SEQ ID NO: 6.
4. A kit for determining whether a sample contains a fungus, the kit comprising: a monoclonal antibody or a scFv that specifically binds the α-1,6 mannose backbone of fungal mannan in said sample, wherein the monoclonal antibody comprises an amino acid sequence encoded by at least a light chain nucleic acid sequence comprising SEQ ID NO: 1 of mAb 2DA6 or at least a heavy chain nucleic acid sequence comprising SEQ ID NO: 2 of mAb 2DA6; and instructional materials for the use thereof.
5. The kit of claim 4 , wherein the monoclonal antibody is mAb 2DA6, and the scFv
scFv clone 11C16 comprising an amino acid sequence encoded by a light chain nucleic acid sequence comprising SEQ ID NO: 5 and a heavy chain nucleic acid sequence comprising SEQ ID NO: 6.
6. A method of making a monoclonal antibody that specifically binds the α-1,6 mannose backbone of fungal mannan, wherein the antibody comprises an amino acid sequence encoded by at least a light chain nucleic acid sequence comprising SEQ ID NO: 1 of mAb 2DA6 or at least a heavy chain nucleic acid sequence comprising SEQ ID NO: 2 of mAb 2DA6, the method comprising:
immunizing a mammal with Saccharomyces Mnn2,
isolating B cells that produce an antibody that specifically binds the α-1,6 mannose backbone of fungal mannan,
fusing the B cells with a cancer cell to generate a hybridoma cell that produces an antibody that specifically binds the α-1,6 mannose backbone of fungal mannan.
7. A method of making a monoclonal antibody that specifically binds the α-1,6 mannose backbone of fungal mannan, wherein the antibody comprises an amino acid sequence encoded by at least a light chain nucleic acid sequence comprising SEQ ID NO: 1 of mAb 2DA6 or at least a heavy chain nucleic acid sequence comprising SEQ ID NO: 2 of mAb 2DA6, the method comprising:
immunizing a mammal with a substance having an effective amount of α-1,6 mannose,
isolating B cells that produce an antibody that specifically binds the α-1,6 mannose backbone of fungal mannan,
fusing the B cells with a cancer cell to make a hybridoma cell that produces an antibody that specifically binds the α-1,6 mannose backbone of fungal mannan.
8. A method of making a monoclonal antibody that specifically binds the α-1,6 mannose backbone of fungal mannan, wherein the antibody comprises an amino acid sequence encoded by at least a light chain nucleic acid sequence comprising SEQ ID NO: 1 of mAb 2DA6 or at least a heavy chain nucleic acid sequence comprising SEQ ID NO: 2 of mAb 2DA6, the method comprising:
immunizing a mammal with a fungus comprising an α-1,6 mannose backbone,
isolating B cells that produce an antibody that specifically binds the α-1,6 mannose backbone of fungal mannan,
fusing the B cells with a cancer cell to make a hybridoma cell that produces an antibody that specifically binds the α-1,6 mannose backbone of fungal mannan.
9. The method of claim 8 , wherein the fungus comprising an α-1,6 mannose backbone is selected from the group consisting of Aspergillus spp., Candida spp., Mucor spp., Rhizopus spp., and Fusarium spp.
10. The method of claim 6 , wherein the mammal that is immunized is a Diversity Outbred mouse.