title: Conformational stability and aggregation of therapeutic monoclonal antibodies studied with ANS and Thioflavin T binding.
abstract: Characterization of aggregation profiles of monoclonal antibodies (mAb) is gaining importance because an increasing number of mAb-based therapeutics are entering clinical studies and gaining marketing approval. To develop a successful formulation, it is imperative to identify the critical biochemical properties of each potential mAb drug candidate. We investigated the conformational change and aggregation of a human IgG1 using external dye-binding experiments with fluorescence spectroscopy and compared the aggregation profiles obtained to the results of size-exclusion chromatography. We show that using an appropriate dye at selected mAb concentration, unfolding or aggregation can be studied. In addition, dye-binding experiments may be used as conventional assays to study therapeutic mAb stability.
authors.affiliation: Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
authors.forename: Veysel
authors.initials: V
authors.lastname: Kayser
authors.forename: Naresh
authors.initials: N
authors.lastname: Chennamsetty
authors.forename: Vladimir
authors.initials: V
authors.lastname: Voynov
authors.forename: Bernhard
authors.initials: B
authors.lastname: Helk
authors.forename: Bernhardt L
authors.initials: BL
authors.lastname: Trout
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000378:metabolism / Q000627:therapeutic use*; D052160:Benzothiazoles; D002850:Chromatography, Gel; D004355:Drug Stability; D005456:Fluorescent Dyes / Q000378:metabolism*; D006801:Humans; D011487:Protein Conformation; D017510:Protein Folding; D055550:Protein Stability; D013050:Spectrometry, Fluorescence; D013696:Temperature; D013844:Thiazoles / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D052160:Benzothiazoles; D005456:Fluorescent Dyes; D013844:Thiazoles; C009462:thioflavin T