title: Oral peptide specific egg antibody to intestinal sodium-dependent phosphate co-transporter-2b is effective at altering phosphate transport in vitro and in vivo.
abstract: Hyperimmunized hens are an effective means of generating large quantities of antigen specific egg antibodies that have use as oral supplements. In this study, we attempted to create a peptide specific antibody that produced outcomes similar to those of the human pharmaceutical, sevelamer HCl, used in the treatment of hyperphosphatemia (a sequela of chronic renal disease). Egg antibodies were generated against 8 different human intestinal sodium-dependent phosphate cotransporter 2b (NaPi2b) peptides, and hNaPi2b peptide egg antibodies were screened for their ability to inhibit phosphate transport in human intestinal Caco-2 cell line. Antibody produced against human peptide sequence TSPSLCWT (anti-h16) was specific for its peptide sequence, and significantly reduced phosphate transport in human Caco-2 cells to 25.3±11.5% of control nonspecific antibody, when compared to nicotinamide, a known inhibitor of phosphate transport (P≤0.05). Antibody was then produced against the mouse-specific peptide h16 counterpart (mouse sequence TSPSYCWT, anti-m16) for further analysis in a murine model. When anti-m16 was fed to mice (1% of diet as dried egg yolk powder), egg yolk immunoglobulin (IgY) was detected using immunohistochemical staining in mouse ileum, and egg anti-m16 IgY colocalized with a commercial goat anti-NaPi2b antibody. The effectiveness of anti-m16 egg antibody in reducing serum phosphate, when compared to sevelamer HCl, was determined in a mouse feeding study. Serum phosphate was reduced 18% (P<0.02) in mice fed anti-m16 (1% as dried egg yolk powder) and 30% (P<0.0001) in mice fed sevelamer HCl (1% of diet) when compared to mice fed nonspecific egg immunoglobulin. The methods described and the findings reported show that oral egg antibodies are useful and easy to prepare reagents for the study and possible treatment of select diseases.
authors.affiliation: Animal Sciences Department, University of Wisconsin-Madison, Madison, WI 53706.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Bobeck
authors.affiliation: Animal Sciences Department, University of Wisconsin-Madison, Madison, WI 53706.
authors.forename: Erica M
authors.initials: EM
authors.lastname: Hellestad
authors.affiliation: Animal Sciences Department, University of Wisconsin-Madison, Madison, WI 53706.
authors.forename: Jordan M
authors.initials: JM
authors.lastname: Sand
authors.affiliation: Animal Sciences Department, University of Wisconsin-Madison, Madison, WI 53706.
authors.forename: Michelle L
authors.initials: ML
authors.lastname: Piccione
authors.affiliation: Animal Sciences Department, University of Wisconsin-Madison, Madison, WI 53706.
authors.forename: Jeff W
authors.initials: JW
authors.lastname: Bishop
authors.affiliation: OPKO Health, Renal Division, Markham, Ontario, Canada L3R 6H3.
authors.forename: Christian
authors.initials: C
authors.lastname: Helvig
authors.affiliation: OPKO Health, Renal Division, Markham, Ontario, Canada L3R 6H3 Department of Biomedical and Molecular Sciences & Cancer Research Institute, Queen's University, Kingston, Ontario, Canada K7L 3N6.
authors.forename: Martin
authors.initials: M
authors.lastname: Petkovich
authors.affiliation: Animal Sciences Department, University of Wisconsin-Madison, Madison, WI 53706 mcook@wisc.edu.
authors.forename: Mark E
authors.initials: ME
authors.lastname: Cook
mesh_terms: D000818:Animals; D000906:Antibodies / Q000378:metabolism; D018938:Caco-2 Cells; D002642:Chick Embryo; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007136:Immunoglobulins / Q000097:blood / Q000235:genetics / Q000276:immunology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010063:Ovum / Q000378:metabolism; D010710:Phosphates / Q000097:blood*; D020539:Sequence Analysis, Protein; D050608:Sodium-Phosphate Cotransporter Proteins / Q000235:genetics* / Q000276:immunology
chemical_list: D000906:Antibodies; C016512:IgY; D007136:Immunoglobulins; D010710:Phosphates; D050608:Sodium-Phosphate Cotransporter Proteins
keywords: IgY; NaPi2b; hen egg antibody; phosphate transport; sevelamer