Affinities of different proteins and peptides for lipopolysaccharide as determined by biosensor technology

Biochem Biophys Res Commun. 1998 Nov 18;252(2):492-6. doi: 10.1006/bbrc.1998.9675.


Biosensor technology was employed to study the specific interactions of different lipopolysaccharide (LPS)-binding proteins and peptides with LPS, using an LPS-coated surface. Two methods to immobilize biotinylated LPS to streptavidin-coated sensor chips (SA-chips) were evaluated. Biotinylated LPS in PBS or biotinylated LPS, pretreated with EDTA and sodium-desoxycholate, were injected across an SA-chip, resulting in a 'high-' and 'low- mass' LPS chip, respectively. While the 'high mass' LPS chip appeared to be unstable, the 'low mass' LPS chip resulted in reproducible binding curves for bactericidal/permeability-increasing protein (rBPI21) with a binding affinity corresponding to the literature (Kd: 3.75 nM). New Kd values were obtained for serum amyloid P component (SAP, Kd: 3.9 nM), a recently discovered new LPS-binding protein, and cationic protein 18 (CAP18, Kd: 0.58 nM). Moreover, binding affinities of bioactive BPI- and SAP-derived peptides could be determined. This study shows for the first time the applicability of biosensor technology to study interactions of proteins and peptides with LPS, using an LPS-coated sensor chip.

MeSH terms

  • Acute-Phase Proteins*
  • Amino Acid Sequence
  • Antimicrobial Cationic Peptides
  • Biosensing Techniques*
  • Blood Proteins / genetics
  • Blood Proteins / metabolism
  • Carrier Proteins / chemistry
  • Carrier Proteins / metabolism
  • Humans
  • In Vitro Techniques
  • Kinetics
  • Lipopolysaccharides / metabolism*
  • Membrane Glycoproteins*
  • Membrane Proteins*
  • Molecular Sequence Data
  • Peptide Fragments / genetics
  • Peptide Fragments / metabolism
  • Peptides / genetics
  • Peptides / metabolism*
  • Protein Binding
  • Proteins / genetics
  • Proteins / metabolism*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Serum Amyloid P-Component / genetics
  • Serum Amyloid P-Component / metabolism
  • Streptavidin


  • Acute-Phase Proteins
  • Antimicrobial Cationic Peptides
  • Blood Proteins
  • Carrier Proteins
  • Lipopolysaccharides
  • Membrane Glycoproteins
  • Membrane Proteins
  • Peptide Fragments
  • Peptides
  • Proteins
  • Recombinant Proteins
  • Serum Amyloid P-Component
  • bactericidal permeability increasing protein
  • lipopolysaccharide-binding protein
  • Streptavidin