In vitro immunoglobulin light chain fibrillogenesis

Methods Enzymol. 1999;309:204-17. doi: 10.1016/s0076-6879(99)09016-3.

Abstract

These techniques permit the production of bulk quantities of fibrils and provide methods for monitoring the kinetics of fibrillogenesis. Experiments performed in the fluorimeter require low protein concentrations, sampling is not necessary (with ThT in situ), and the measured fluorescence signal is indicative of fibril content and is not complicated by the presence of amorphous aggregates. However, ASF using the orbital shaker is a simple, rapid, initial procedure, adequate for screening for fibrillogenic potential, in which multiple experiments can be performed simultaneously and over long periods of incubation. These methods may be used to investigate the fibrillogenesis of VL proteins and BJps as a means of predicting pathogenicity, as well as providing information on the basic biophysical principles underlying light chain aggregation.

MeSH terms

  • Amino Acid Sequence
  • Amyloid / metabolism*
  • Bence Jones Protein / metabolism
  • Benzothiazoles
  • Humans
  • Immunoglobulin Light Chains / genetics
  • Immunoglobulin Light Chains / metabolism*
  • Molecular Sequence Data
  • Recombinant Proteins / metabolism
  • Thiazoles / analysis
  • Thiazoles / metabolism

Substances

  • Amyloid
  • Benzothiazoles
  • Immunoglobulin Light Chains
  • Recombinant Proteins
  • Thiazoles
  • thioflavin T
  • Bence Jones Protein