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Clinical Chemistry
Protein and electrophoresis
| Question | Answer |
|---|---|
| has both + and - regions | zwitteron |
| The carboxylic end on one amino acid is attached to the amine group on another amino acid by a _______ bond | peptide |
| tertiary structure | 3D shape (domain) |
| Digestion involves ___ and ____ from the stomach and _____ and _________ from the pancreas | HCl & pepsin// Trypsin & chymotrypsin |
| Amino acids absorbed thru intestinal mucosa into ___________ circulation; go to liver | hepatic portal |
| synthesis of new proteins is in the ______ | liver |
| ___ proteins leave in urine | small molecular weight |
| Protein functions | -maintain osmotic pressure -buffer capacity by Hb and plasma proteins -transport molecules - structural support |
| structural support proteins | -collagen -keratin |
| transport proteins | -Albumin -transferrin -apoproteins -ceruloplasmins |
| ________ regulates fluid movement between intravascular & interstitial compartments | Albumin |
| OTHER functions of proteins | 2nd messengers, enzymes, coagulation factors, antibodies, contractile myofibrils |
| 2 kinds of contractile myofibrils | actin & myosin |
| Albumin accounts for __ of all plasma proteins | 60% |
| Functions of Albumin | -maintain osmotic pressure -transports substances in blood |
| Albumin serum reference range | 3.5-5.5 g/dL |
| indicator of nutritional status | Pre-albumin |
| Causes of increased serum albumin | -Dehydration, fluid loss (relative increase) -Sunstroke, strenuous exercise -Multiple sclerosis -Hypothyroidism |
| Decreased serum albumin | -Malnutrition, malabsorption of proteins -Liver disease – ineffective synthesis by hepatocytes -Kidney disease – loss in urine from glomerular -damage -Burns, pregnancy, chronic infections |
| most common binding method | colorimetric assay using HABA, brom green/purple |
| Healthy people excrete ___ mg/day of albumin | 30 |
| Macroalbumin | >300mg/day |
| microalbumin | 30-300mg/day |
| globulin proteins that are elevated with inflammation | Alpha-1 |
| binds free hemoglobin | haptoglobin (alpha-2) |
| types of globulin proteins | -Alpha 1 -Alpha 2 -Beta -Gamma |
| hereditary disease that affects liver and lungs | Alpha-1-Antitrypsin (A1AT) Deficiency |
| People with Alpha-1-Antitrypsin lack ___ | alpha 1 fraction |
| most common case of liver disease in kids | alpha-1-antitrypsin |
| Correlation of markers for infection & inflammation | increasd levels of: -Alpha-1-antitrypsin -C3 & C4 (complement fractions) -CRP (C-reactive protein) |
| protein markers related to anemia | -transferrin -haptoglobin -ferritin |
| increased in iron deficiency anemia (IDA) | Transferrin |
| Decreased in hemolysis | Haptoglobin |
| Decreased in IDA | Ferritin |
| Reference range for serum total proteins | 6.5-8.3g/dL |
| cause of elevated total protein | -fluid loss -increase in gobulin fraction, usually gamma |
| cause of decrease total protein | -protein loss -dec. protein synthesis -dec. dietary protein intake |
| Reference method; measures nitrogen content after digestion of protein; very laborious | Kjedahl |
| Compares refractive index due to dissolved solutes with RI of pure water; rapid, simple, cheap | refractometry |
| Routine, colorimetric method; violet color proportional to # peptide bonds in proteins | Biuret |
| Glomerular damage, kidney tubule dysfunction, exceed renal threshold for protein (Bence Jones) causes | increased urine protein |
| CNS infection, obstruction, neoplasm, trauma; cerebral infarction; multiple sclerosis | causes of increased CSF protein |
| -Protein precipitated with dilute acid (SSA, TCA) -amount of turbidity proportional to protein | Turbidimetric |
| positively charged end | anode |
| negatively charged end | cathode |
| Separation of proteins in an electric field based on net charge | electrophoresis |
| determines pH; higher concentration means slower migration | buffer concentration |
| Neutral proteins (gamma globulins) migrate toward cathode band displacement | electroendosmosis |
| higher voltage = | faster migration |
| protein that migrates fastest toward anode | albumin |
| Albumin is towards the ____ side on the electrophoresis gel | + anode |
| Technique to characterize monoclonal proteins -Couples electrophoresis with immunochemical reactions in agarose | IMMUNOFIXATION ELECTROPHORESIS (IFE) |
| Urine Electrophoresis: Must be concentrated ____ fold before application | 50-100 |
| Urine Electrophoresis: Can correlate serum & urine electrophoresis patterns for diseases like _____ | multiple myeloma |
| CSF Electrophoresis: needs to be concentrated ___ fold | 50x |
| CSF Electrophoresis: Used to help identify abnormal “_____” bands in gamma region in ______. Many persons with this disease have increased synthesis of _______ in their CSF | oligoclonal// multiple sclerosis// immunoglobulins |
| Calculation of ___;___ ratio can help diagnose MS | gamma globulin: albumin ratio |
| To determine if elevated CSF IgG is due to local synthesis or leakage from plasma, can calculate IgG index | CSF IgG x serum albumin/ serum IgG x CSF albumin |
| Normal CSF igG index | 0.26-0.70 |
| Not done frequently since quantitation of individual lipoproteins (HDL, LDL, VLDL) can be done on automated chem analyzers in less time | LIPOPROTEIN ELECTROPHORESIS |
| LIPOPROTEIN ELECTROPHORESIS: After electrophoretic separation, lipoprotein fractions stained with fat-soluble dye such as ____ -Quantitate in scanning_________ | fat red 7B// densitiometer |
| Often associated with a malignant process | Monoclonal Gammpathies |
| Monoclonal Gammopathies: Overproduction of 1 clone of ______, causing a monoclonal (M) “spike” in gamma region on protein electrophoresis -Serum total protein & gamma globulins increased | immunoglobulin |
| Examples of Monoclonal Gammopathy | Multiple myeloma// Waldenstom's macroglobulinemia |
| Proliferation of many clones of plasma cells, so total gamma globulin level increased | Polyclonal gammopathy |
| increase of all globulins | chronic infalmmation |
| : increase of gamma globulins | Hepatitis |
| beta-gamma bridging | liver cirrhosis |
Created by:
lindsayburch