Label-Free, Direct Measurement of Protein Concentrations in Turbid Solutions with a UV-Visible Integrating Cavity Absorbance Spectrometer

Anal Chem. 2018 Apr 17;90(8):4982-4986. doi: 10.1021/acs.analchem.8b00502. Epub 2018 Mar 30.

Abstract

Protein-particle conjugates and mixtures have been investigated extensively for their diverse applications in biotechnology. However, general methods to measure protein concentration of protein-particle solutions are lacking. Typically, proteins in turbid solutions require separation or staining with another chromophore to quantitate their concentration. Here we demonstrate a label-free, direct approach to measure protein concentrations in turbid solutions using a UV-vis integrating cavity absorbance spectrometer. Three systems are used to test the ability to measure accurate protein concentrations: proteins adsorbed to Alhydrogel, proteins in solution with gold nanoparticles, and proteins encapsulated within polymeric microspheres. Protein concentrations in each of the three protein-particle systems were successfully quantified using a calibration curve created from the absorbance at 280 nm.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aluminum Hydroxide / chemistry
  • Animals
  • Calibration
  • Cattle
  • Gold / chemistry
  • Immunoglobulin G / analysis
  • Metal Nanoparticles / chemistry
  • Polylactic Acid-Polyglycolic Acid Copolymer / chemistry
  • Proteins / analysis*
  • Proteins / standards
  • Serum Albumin, Bovine / analysis
  • Serum Albumin, Bovine / standards
  • Solutions / chemistry*
  • Spectrophotometry* / standards

Substances

  • Immunoglobulin G
  • Proteins
  • Solutions
  • Polylactic Acid-Polyglycolic Acid Copolymer
  • Serum Albumin, Bovine
  • Aluminum Hydroxide
  • Gold