Presentation on theme: "Serum Protein Electrophoresis"— Presentation transcript:
1Serum Protein Electrophoresis Dr. Nitin InamdarDepartment of BiochemistryTata Memorial Center, Parel,Mumbai.
2Electrophoresis Phoresis: Separation or migration or movement Electro: Under influence of electric field
3Constant 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
4Electrophoresis 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.
5Serum Electrophoresis Paper electrophoresisStarch gel electrophoresisAgarosre gel electrophoresisCellulose acetate gel electrophoresisCapillary gel electrophoresis
6Serum protein electrophoresis on agarose gel is a type of horizontal gel electrophoresis
7Equipment used for the gel electrophoresis power supply (direct current)containers for staining and destaining gelelectrophoresis chamberapplicator
16WHO 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
17Diagnostic 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
18Immunoglobin (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.
39NephelometryAntigen-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.
40Immunofixation 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
53light 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.
54Freelite 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