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Dr. Nitin Inamdar Department of Biochemistry Tata Memorial Center, Parel, Mumbai.

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Presentation on theme: "Dr. Nitin Inamdar Department of Biochemistry Tata Memorial Center, Parel, Mumbai."— Presentation transcript:

1 Dr. Nitin Inamdar Department of Biochemistry Tata Memorial Center, Parel, Mumbai.

2  Phoresis: Separation or migration or movement  Electro: Under influence of electric field

3 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

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) and the size and shape of the subsets of these proteins are used in interpreting the results.

5 Paper electrophoresis Starch gel electrophoresis Agarosre gel electrophoresis Cellulose acetate gel electrophoresis Capillary gel electrophoresis

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7 power supply (direct current) electrophoresis chamber containers for staining and destaining gel applicator

8  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  Diagnostic: ◦ Diagnosis of plasma cell dyscrasia ◦ Diagnosis of Waldenstrom’ macroglbulinemia  Monitoring of disease: ◦ Monitoring of plasma cell dyscrasia

10 1. Serum protein electrophoresis 2. Quantitative immunoglobulins (nephelometry) 3. Immunofixation, immunoelectrophoresis and immunodiffusion 4. Urine studies 5. light chain

11  Albumin  Globulins: ◦ α1, ◦ α2, ◦ β1, ◦ β2, ◦ gamma  A/G Ratio: 2.0

12 Normal serum electrophoresis pH=8.6, 1% agarose gel, sodium barbitone buffer Anode +veCathode -ve

13 Abnormal serum electrophoresis showing M band Anode +veCathode -ve

14 Normal serum electrophoresis

15 Abnormal serum electrophoresis showing M band

16  Plasma cell myeloma (MM)  Plasma cell myeloma variants Non secretory myeloma Indolent Myeloma Smoldering Myeloma Plasma cell leukemia  Plasmacytomas Solitary plasmacytoma of bone Extramedullary plasmacytoma  Immunoglobulin deposition diseases Primary Amyloidosis Systemic light and heavy chain deposition disease  Osteosclerotic myeloma (POEMS syndrome)  Heavy chain disease Gamma /Mu / Alpha

17 Symptomatic plasma cell myeloma: BM clonal plasma cells or Plasmacytoma Marrow 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/dL Marrow Plasmacytosis < 10% No Lytic bone lesions or related organ impairment etc

18 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.

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39  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.

40  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

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53 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

54 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 Freelite test

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