Presentation on theme: "Serum Protein Electrophoresis"— Presentation transcript:
1 Serum Protein Electrophoresis Dr. Nitin InamdarDepartment of BiochemistryTata Memorial Center, Parel,Mumbai.
2 Electrophoresis Phoresis: Separation or migration or movement Electro: Under influence of electric field
3 Constant current 5mAM/Slide SerumElectrophoresisMixture of proteinsAmino 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 buffer1%AgaroseConstant current 5mAM/Slide
4 Electrophoresis separates proteins based on their physical properties.Serum is placed on a specific medium, and a charge is applied.The net charge (positive or negative) andthe size and shape of the subsets of theseproteins are used in interpreting the results.
5 Serum Electrophoresis Paper electrophoresisStarch gel electrophoresisAgarosre gel electrophoresisCellulose acetate gel electrophoresisCapillary gel electrophoresis
6 Serum protein electrophoresis on agarose gel is a type of horizontal gel electrophoresis
7 Equipment used for the gel electrophoresis power supply (direct current)containers for staining and destaining gelelectrophoresis chamberapplicator
8 Various electrophoresis 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
9 Indication of serum electrophoresis Diagnostic:Diagnosis of plasma cell dyscrasiaDiagnosis of Waldenstrom’ macroglbulinemiaMonitoring of disease:Monitoring of plasma cell dyscrasia
10 Method of detection Serum protein electrophoresis Quantitative immunoglobulins (nephelometry)Immunofixation, immunoelectrophoresis and immunodiffusionUrine studieslight chain
16 WHO Classification of Plasma Cell Neoplasms Plasma cell myeloma (MM)Plasma cell myeloma variantsNon secretory myelomaIndolent MyelomaSmoldering MyelomaPlasma cell leukemiaPlasmacytomasSolitary plasmacytoma of boneExtramedullary plasmacytomaImmunoglobulin deposition diseasesPrimary AmyloidosisSystemic light and heavy chain deposition diseaseOsteosclerotic myeloma (POEMS syndrome)Heavy chain diseaseGamma /Mu / Alpha
17 Diagnostic criteria for plasma cell dyscrasia Symptomatic plasma cell myeloma:BM clonal plasma cells or PlasmacytomaMarrow plasmacytosis (>30%)M-component in serum or urineSerum: Ig G >3 g/dL, Ig A >2.5 g/dL orUrine: >/= 1g/ day of λ or κ [BJP]Asymptomatic plasma cell myeloma:M band in serum at myeloma levels > 3 gm/dLand /or10% or more clonal plasma cell in BMNo related organ or tissue impairment (end organ damage or bony lesion) or myeloma-related symptomsMUGS:M-Component, present but less than 3 gm/dLMarrow Plasmacytosis < 10%No Lytic bone lesions or related organ impairment etc
18 Immunoglobin (Ig) structure and types Malignant Plasma Cell (Clonal Proliferation) produces one type ofIg either A,M, G, D or E with kappa or lambda light chains in excess amount.
39 NephelometryAntigen-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.
40 Immunofixation electrophoresis 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
53 light chain myelomaA 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.
54 Freelite test Free Kappa Free Lambda Kappa/ Lambda ratio Normal serum mean median %concentration concentration rangeFree Kappa8,36 mg/l ,30 mg/l ,30 – 19,40 mg/lFree Lambda13,43 mg/l ,40 mg/l ,71 – 26,30 mg/lKappa/Lambda ratiomean median total range0, , ,26 – 1,65