12. Laboratory analysis of plasma proteins

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Plasma proteins

  • Total protein
    • Normal serum level 66 - 83 g/L
    • Normal urine level < 0,1 g/L
    • Biuret test (copper forms purple complex with proteins in alkaline solution)
    • Colour intensity proportional to protein concentration – photometry
  • Albumin
    • Normal serum level 35 - 53 g/L
    • Albumin forms green complex with BCG at pH 4,1
    • Photometry
  • Specific proteins
    • Measurement based on immunogenicity – use of monoclonal or polyclonal antibodies
    • Immunoassay – biochemical test measuring concentration of protein by using antibody
      • Enzyme immunoassay (EIA)
      • Enzyme-linked immunosorbent assay (ELISA)
      • Radioimmunoassay (RIA)
      • Fluorescence
    • Proteins
      • Microalbumin (high sensitivity albumin measurement)
      • Hormones
      • Tumour markers
      • Drugs
      • B12
      • Folic acid
      • Transferrin
      • Ferritin
      • EPO
      • HCG
      • Troponin
      • Procalcitonin
  • Separation techniques
    • Chromatography
    • Electrophoresis – sample added to gel -> electric field applied -> molecules separated by electric charge -> stained to make charged molecules visible
      • Qualitative, semi-quantitative!
      • Serum, not plasma (fibrinogen forms a band)
      • Serum electrophoresis – monoclonal paraproteins
      • CSF electrophoresis – multiple sclerosis
      • Haemoglobin electrophoresis – haemoglobinopathies
      • Lipid electrophoresis – dyslipidaemias
      • Urine electrophoresis – Bence-Jones protein

Serum electrophoresis

  • Serum electrophoresis
    • Prealbumin=transthyretin
    • Albumin
    • Alpha-1 band
      • alpha1-antitrypsin
      • alpha1-acid glycoprotein (AGP)
      • alpha1-microglobulin
    • Alpha-2 band
      • Haptoglobin
      • alpha2-macroglobulin
      • Caeruloplasmin
    • Beta band
      • Transferrin
      • Beta2-microglobulin
      • Complement factors
    • Between beta and gamma
      • CRP
    • Gamma band
      • Immunoglobulins