title: Methods for the detection and serum depletion of porcine galectin-3.
abstract: BACKGROUND
  Circulating galectin-3 (Gal-3) is elevated in systemic inflammatory disorders, fibrotic diseases, and in cancers. Gal-3 is a promising cancer target where it promotes tumorigenesis and metastasis, as well as in renal, pulmonary, hepatic, and cardiovascular diseases, because of its role as a driver of fibrotic remodeling. This reports goal was to establish methods for the detection and removal of porcine Gal-3 that will enable further studies of the therapeutic potential of Gal-3 depletion by apheresis in porcine disease models. The long-term aim is to develop a safe, effective method of removing Gal-3 via apheresis as a standalone therapeutic tool and as an adjuvant to other therapies.
  
  
  METHODS
  Purified recombinant porcine Gal-3 was prepared and used as the standard for development of a porcine Gal-3 enzyme-linked immunosorbent assay (ELISA). Different affinity column matrices that incorporated either a rat IgG2a anti-Gal-3 monoclonal antibody or carbohydrate ligand were assessed for depletion of Gal-3 from porcine serum.
  
  
  RESULTS
  A porcine Gal-3 ELISA with a linear range from 0.3 to 20 ng/mL was able to detect native porcine Gal-3 in both fetal (∼150-200 ng/mL) and juvenile (∼5-15 ng/mL) porcine serum samples. Use of an anti-Gal-3 monoclonal antibody affinity column depleted Gal-3 from porcine serum to at least 313 pg/mL, the limit of ELISA detection.
  
  
  CONCLUSIONS
  Methods have been developed for the detection and depletion of porcine Gal-3. These methods will be used to study the specific effects of Gal-3 depletion via apheresis in porcine models of disease.
authors.affiliation: Eliaz Therapeutics Inc, Santa Rosa, California.
authors.forename: Isaac
authors.initials: I
authors.lastname: Eliaz
authors.affiliation: Center for Transplantation Sciences, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.
authors.forename: Aarti
authors.initials: A
authors.lastname: Patil
authors.affiliation: Center for Transplantation Sciences, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.
authors.forename: Nalu
authors.initials: N
authors.lastname: Navarro-Alvarez
authors.affiliation: Center for Transplantation Sciences, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.
authors.forename: Zhirui
authors.initials: Z
authors.lastname: Wang
authors.affiliation: University of California, Los Angeles, California.
authors.forename: Amity
authors.initials: A
authors.lastname: Eliaz
authors.affiliation: Eliaz Therapeutics Inc, Santa Rosa, California.
authors.forename: Elaine
authors.initials: E
authors.lastname: Weil
authors.affiliation: Eliaz Therapeutics Inc, Santa Rosa, California.
authors.forename: Barry
authors.initials: B
authors.lastname: Wilk
authors.affiliation: Center for Transplantation Sciences, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.
authors.forename: David H
authors.initials: DH
authors.lastname: Sachs
authors.affiliation: Center for Transplantation Sciences, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.
authors.forename: Christene A
authors.initials: CA
authors.lastname: Huang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D000918:Antibody Specificity; D001781:Blood Component Removal / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D037502:Galectin 3 / Q000097:blood* / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D007074:Immunoglobulin G; D051381:Rats; D011994:Recombinant Proteins / Q000097:blood / Q000276:immunology / Q000302:isolation & purification; D013552:Swine
chemical_list: D000911:Antibodies, Monoclonal; D037502:Galectin 3; D007074:Immunoglobulin G; D011994:Recombinant Proteins
keywords: ELISA; affinity depletion; galectin-3; porcine