Binding moieties for biofilm remediation
Binding agents able to disrupt bacterial biofilms of diverse origin are described, including monoclonal antibodies secreted by human B lymphocytes. Methods to prevent formation of or to dissolve biofilms with these binding agents are also described. Immunogens for eliciting antibodies to disrupt biofilms are also described.
1. A method to obtain antibodies immunoreactive with integration host factor (IHF) protein or B cells that secrete antibodies immunoreactive with IHF protein which method comprises assessing whether an antibody present in a patient sample binds to a peptide that consists of the amino acid sequence IEYLSDKYHLSKQDTK (SEQ ID NO:49) (positions 10-25 of the Haemophilus influenzae IHFα chain) or the amino acid sequence RDKSSRPGRNPKTGDVVAARSARR (SEQ ID NO:50) (positions 56-78 of the Haemophilus influenzae IHFα chain), or homologs thereof, and isolating an antibody that binds to the peptide.
2. The method of claim 1 , further comprising testing the affinity of the antibody for a DNABII protein over a range of pH conditions.
3. The method of claim 2 , wherein the pH range comprises a pH at 4.5.
4. The method of claim 1 , further comprising comparing the affinity between the isolated antibodies and a DNABII protein versus the affinity between a component of a biofilm and the DNABII protein.
5. The method of claim 4 , wherein the component of the biofilm is a branched DNA.
6. The method of claim 4 , wherein the comparing step is performed as a competitive assay.
7. The method of claim 4 , wherein the comparing step comprises determining the affinity independently.
8. The method of claim 1 , wherein the assessing step comprises assessing whether the antibody compete for the monoclonal antibody having a variable region which comprises the heavy and light chain variable regions of TRL1068 (SEQ ID NO:1) and (SEQ ID NO:2) or of TRL1330 (SEQ ID NO:21) and (SEQ ID NO:22) or of TRL1337 (SEQ ID NO:25) and (SEQ ID NO:26).
9. The method of claim 1 , further comprising administering the peptide to a subject.
10. The method of claim 9 , further comprising sequencing the antibody.
11. The method of claim 9 , further comprising recovering B cells from the subject.