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33 Cards in this Set
- Front
- Back
endosmosis? which proteins are weakly anionic and swept up in end osmotic flow?
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electrophoresis:
**a charge is applied across a medium that is composed of a solid support and a fluid buffer. **this charge creates an electromotive force **the solid support has a slight negative charge and is drawn towards the positive pole (anode), but solid support cannot move . **There is compensatory flow of the fluid buffer towards the negative pole (cathode) . This buffer flow is called endosmosis and has the capacity to carry with it substances suspended within the medium . *If proteins are added to the medium and the charge is applied, two forces act on each protein : electromotive and endosmotic; electromotive force tends to pull them towards the anode (positive pole) ; whereas, endosmosis pulls them towards the cathode . **In the case of gamma globulins, which have a weak net negative charge, the endosmotic force displaces the proteins towards the cathode . In other proteins,electromotive force exceeds the endosmotic force, and they move to anode |
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what eletrophoretic technique resolves various forms of a single protein and is used in clinical laboratories?
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isoelectric focusing; SDS-PAGE is an alternative method, but it is used chiefly in research laboratories
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most common analyze measured by affinity chromatography? what chemical is used to bind the molecule of interest?
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quantization of glycosylated hemoglobin; idihydroxyboronate
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drop of serum is placed on a refractometer
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specific gravity
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acid digestion releases ammonium ions which are then quantitated using iodine
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kjeldahl technique
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copper salts in alkaline solution form a purple complex with substances with two or more peptide bonds
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biuret method
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most common dye-binding method to quantitate albumin
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bromcresol green
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used often to detect peptides after paper chromatography or ion exchange
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ninhydrin
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an acid such as tricholoroacetic forms a precipitate which clouds the solution
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turbidimetry
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chief physiologic function of pre albumin?
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binds vitamin A along with retinol-binding protein to transport vitamin A throughout the body
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clinical significance of a pre albumin band in CSF?
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normal findings; as a compact molecule prealbumin crosses the CSF barrier more readily than do other proteins
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do patents with analbuminemia suffer from as cities and edema, as do cirrhotics?
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no; the patient has compensatory mechanisms to maintain oncotic pressure
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A, Serum protein electrophoresis in cirrhosis (Cir) shows more rapidly migrating albumin compared with normal serum (Nor) because of theadditional negative charge from covalently linked conjugated bilirubin (i.e., δ-bilirubin). The γ-globulins are broadly increased in the polyclonal elevation char-acteristic of cirrhosis of the liver. B, Protein electrophoretic patterns of serum (Ser) and concentrated urine (Ur) in a patient with nephrotic syndrome. Smaller molecular-sized proteins such as albumin are preferentially lost from blood into urine. Larger proteins such as α2-macroglobulin and β-lipoprotein are retained in the blood and constitute major bands in the serum pattern.
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Immunofixation with transferrin antibody to test for presence of cerebrospinal fluid (CSF). Ser, Normal serum showing position of transferrin (Tf; double-headed arrow); CSF, normal position of Tf and asialotransferrin (aTf, single-headed arrow); Pt Ser, patient serum included to rule out electrophoretic variant ofTf; Pt Fl, unknown fluid from patient demonstrating bands of both Tf and aTf that confirm the presence of CSF in that fluid.
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23 yo pregnant woman with an iron of 27microgram/dL and iron binding capacity of 603microgram/dL; increased transferrin
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Serum protein patterns in (1) chronic inflammation with decreased albumin and increased γ-globulins; (2) acute inflammation with increased α2-fraction (haptoglobin) and decreased C3 due to activation and consumption of complement; (3) inanition post–spinal cord injury with hypoproteinemia of several fractions.
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1. 28yom HIV with marked peripheral edema; Glomerular proteinuria, transferrin
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Patterns of urine protein electrophoresis in different disorders. (1) Severe glomerular proteinuria with a major band of albumin plus a secondary one of transferrin (*). (2) Trace proteinuria with a faint band of albumin and other diffuse proteins.
(3) Immunoglobulin light chains (*). (4) Tubular proteinuria with multiple bands that do not correspond to major serum proteins. (5) Hematuria with a majorband of hemoglobin (not to be confused with monoclonal gammopathy), in addi-tion to albumin. |
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Serum and urine protein electrophoretic patterns in a patient with multiple myeloma. Serum demonstrates a predominance of the larger complete immunoglobulin; the urine has a large amount of the smaller-sized light chains with only a small amount of the whole immunoglobulin.
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Decreased albumin and variable decreases in all globulins except a-2 macroglobulin, consistent with nephrotic syndrome. May cause a "pseudo M~protein " No restriction band on IFE
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IgM K M-protein . Note that IgM paraproteins tend to migrate nearer beta
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IgA ~, M-protein . Note that IgA paraproteins tend to migrate into the beta region.
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IgG kappa M-protein
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Acute inflammation . Decreased albumin with increased a-1 & a-2 globulins
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Chronic inflammation: mildly decreased albumin, increased a-1, a-2, and gamma globulins
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Marked changes of chronic inflammation, beta-gamma bridging, (and no restriction band on IFE) in a patient with primary biliary cirrhosis (PBC)
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CSF
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UPEP
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What acute phase reactant rises most quickly and returns to normal most quickly in inflammatory states?
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C-reactive protein
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What methods are used to screen patients suspected of having the ZZ phenotype of AAT deficiency?
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Both nephelometry and serum protein electrophoresis are used in screening; confirmation must be performed with ancillary tests; the ZZ phenotype shows marked decrease levels of AAT
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What disease state is alpha 2 macroglobulin most increased?
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Nephrotic syndrome
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Does haptoglobin bind myoglobin in cases of rhabdomyolysis?
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No; haptoglobin scavengers hemoglobin in times of hemolysis but not myoglobin
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17 yo admitted after MVA; how would you determine the clear fluid draining from his left nasal passage?
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Transferrin
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38 yof on oral contraceptives being treated with heparin for pulmonary embolism has blood drawn in a red top tube serum protein electrophoresis (SPEP); a distinct band noted just anodal to the gamma fraction?
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Band is fibrinogen; heparin iced patients' blood does not completely clot
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