Antibodies and methods for generating the same
OwnerAndrew PORTER, Soumya PALLIYIL +1inventors
InventorsAndrew PORTER, Soumya PALLIYIL, Carol MUNRO
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Abstract
Provided are methods for identifying surface-exposed epitopes of fungal cell wall proteins, and related peptide antigens, suitable for the development of antifungal antibodies. The methods allow for the detection of surface-exposed epitopes that are particularly highly expressed in drug-resistant fungal pathogens, and in pathogens exposed to antifungal drugs. Also provided are antifungal antibodies which may be derived by these methods. The methods and antibodies find use in the treatment of treating and diagnosing fungal infections, such as candidiasis, aspergillosis cryptococcosis, and Mucormycosis.
Claims30
A method for identifying epitopes of fungal cell wall proteins (“CWPs”) suitable for the development of antifungal antibodies, comprising:
- (a) providing a population of a first strain of a fungal species;
- (b) culturing:
- (i) a first sub-population of the first strain in the presence of an antifungal agent, thereby forming a first culture;
- (ii) a second sub-population of the first strain in the absence of an antifungal agent, thereby forming a second culture;
- (c) harvesting the cells and isolating the cell walls from each culture;
- (d) digesting the cell walls from each culture with a digesting agent for surface-exposed CWPs, thereby forming digested surface-exposed fungal CWPs;
- (e) performing proteomic analysis on the digested surface-exposed fungal CWPs;
- and
- (f) identifying surface-exposed epitopes of the digested surface-exposed CWPs that are expressed in higher abundance in the first culture compared to the second culture.
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A method for identifying epitopes of fungal cell wall proteins (“CWPs”) suitable for the development of antifungal antibodies, comprising:
- (a) providing populations of a first strain and a second strain of a fungal species, wherein the first strain is resistant to an antifungal drug, and the second strain is not resistant to an antifungal drug;
- (b) culturing:
- (i) a first sub-population of the first strain in the presence of an antifungal agent, thereby forming a first culture;
- (ii) a second sub-population of the first strain in the absence of an antifungal agent, thereby forming a second culture;
- (iii) a first sub-population of the second strain in the presence of an antifungal agent, thereby forming a third culture;
- and
- (iv) a second sub-population of the second strain in the absence of an antifungal agent, thereby forming a fourth culture;
- (c) harvesting the cells and isolating the cell walls from each culture;
- (d) digesting the cell walls from each culture with a digesting agent for surface-exposed CWPs, thereby forming digested surface-exposed fungal CWPs;
- (e) performing proteomic analysis on the digested surface-exposed fungal CWPs;
- and
- (f) identifying surface-exposed epitopes of the digested surface-exposed CWPs that are:
- (i) expressed in higher abundance in the first culture compared to the second culture;
- and/or
- (ii) expressed in higher abundance in the third culture compared to the fourth culture;
- and/or
- (iii) expressed in higher abundance in the first culture compared to the third culture.
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An antibody, or a polynucleotide encoding the same, wherein the antibody specifically binds to a surface-exposed cell wall protein (“CWP”), wherein the CWP is Pga31, Utr2, Phr2, or Cht2.
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Dependencies are inferred from the claim text.