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TMH proceedings 2010-2011,pdf
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Serum Protein Electrophoresis
Dr. Nitin Inamdar Department of BiochemistryTata Memorial Center, Parel,Mumbai.
Phoresis: Separation or migration or movement
Electro: Under influence of electric field
Electrophoresis
Serum
Electrophoresis
Mixture of proteins
Amino acids:◦ Amino group (NH2)◦ Carboxyl group (COOH)
Two amino acids join each other with polypeptide bond to form polypeptide chain & many polypeptide chains make a protein.
Ph 8.6/0.075M Barbitone buffer
1%Agarose
Constant current 5mAM/Slide
Electrophoresis separates proteins based ontheir physical properties.
Serum is placed on a specific medium, and a charge is applied. The net charge (positive or negative) and
the size and shape of the subsets of these proteins are used in interpreting the results.
Serum ElectrophoresisPaper electrophoresis
Starch gel electrophoresisAgarosre gel electrophoresis
Cellulose acetate gel electrophoresisCapillary gel electrophoresis
Equipment used for the gel electrophoresis
power supply (direct current)
electrophoresis chamber
containers for staining and destaining gel
applicator
Serum/protein electrophoresis: ◦ plasma cell dyscrasia, nephrotic syndrome, etc.
Hemoglobin electrophoresis: ◦ thalassemia, hemoglobinopathies like sickle cell
anemia, HbC, HbD, etc.
Urine electrophoresis: ◦ BJP in plasma cell dyscrasia
Various electrophoresis
Diagnostic: ◦ Diagnosis of plasma cell dyscrasia◦ Diagnosis of Waldenstrom’ macroglbulinemia
Monitoring of disease: ◦ Monitoring of plasma cell dyscrasia
Indication of serum electrophoresis
1. Serum protein electrophoresis2. Quantitative immunoglobulins
(nephelometry)3. Immunofixation, immunoelectrophoresis
and immunodiffusion4. Urine studies5. light chain
Method of detection
Plasma cell myeloma (MM) Plasma cell myeloma variants
Non secretory myelomaIndolent MyelomaSmoldering MyelomaPlasma cell leukemia
PlasmacytomasSolitary plasmacytoma of boneExtramedullary plasmacytoma
Immunoglobulin deposition diseasesPrimary AmyloidosisSystemic light and heavy chain
deposition disease Osteosclerotic myeloma (POEMS
syndrome) Heavy chain disease
Gamma /Mu / Alpha
WHO Classification of Plasma Cell Neoplasms
Symptomatic plasma cell myeloma:BM clonal plasma cells or PlasmacytomaMarrow plasmacytosis (>30%)M-component in serum or urine
Serum: Ig G >3 g/dL, Ig A >2.5 g/dL or Urine: >/= 1g/ day of λ or κ [BJP]
Asymptomatic plasma cell myeloma:M band in serum at myeloma levels > 3 gm/dL and /or 10% or more clonal plasma cell in BM No related organ or tissue impairment (end organ
damage or bony lesion) or myeloma-related symptoms MUGS:
M-Component, present but less than 3 gm/dLMarrow Plasmacytosis < 10%No Lytic bone lesions or related organ impairment etc
Diagnostic criteria for plasma cell dyscrasia
Immunoglobin (Ig) structure and types
Malignant Plasma Cell (Clonal Proliferation) produces one type of Ig either A,M, G, D or E with kappa or lambda light chains in excess amount.
Nephelometry Antigen-antibody complexes, when formed at a
high rate, will precipitate out of a solution resulting in a turbid or cloudy appearance.
• Nephelometry: is indirect measurement of light scattered by the antigen-antibody complexes.
After separation of serum proteins by electrophoresis separated serum in different lanes is incubated with monospecific anti sera against IgA, M G and κ λ
Immunofixation electrophoresis
A type of multiple myeloma in which plasma cell tumors produce only monoclonal light chain proteins. Persons with light chain disease may develop lytic bone lesions, hypercalcemia, impaired kidney function, and amyloidosis.
light chain myeloma
Normal serum mean median 95 %concentration concentration range
Free Kappa 8,36 mg/l 7,30 mg/l 3,30 – 19,40 mg/l
Free Lambda 13,43 mg/l 12,40 mg/l 5,71 – 26,30 mg/l
Kappa/
Lambda ratio
mean median total range
0,63 0,60 0,26 – 1,65
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