177:
173:
electric current is applied. Proteins with a negative charge will migrate towards the positively charged anode. Albumin has the most negative charge, and will migrate furthest towards the anode. Endoosmotic flow is the movement of liquid towards the cathode, which causes proteins with a weaker charge to move backwards from the application site. Gamma proteins are primarily separated by endoosmotic forces. The drawing of the electrophoretic bands provided by the laboratory may be difficult to remember, and medical students, residents, nurses, and non-specialized medical practitioners may find visual mnemonics useful to recall the five main bands and the shape of normal serum electrophoresis.
199:
29:
576:(mAb), also migrate in this region and may be misinterpreted as a monoclonal gammopathy, and may also be identified by immunofixation or immunodisplacement/immunosubtraction as they are structurally comparable to human immunoglobulins. The most common cause of a restricted band is an MGUS (monoclonal gammopathy of uncertain significance), which, although a necessary precursor, only rarely progresses to multiple myeloma. (On average, 1%/year.) Typically, a monoclonal gammopathy is malignant or clonal in origin,
1380:
1392:
66:
404:. Due to its large size, AMG cannot pass through glomeruli, while other lower-molecular weight proteins are lost. Enhanced synthesis of AMG accounts for its absolute increase in nephrotic syndrome. Increased AMG is also noted in rats with no albumin indicating that this is a response to low albumin rather than nephrotic syndrome itself
172:
Proteins are separated by both electrical forces and electroendoosmostic forces. The net charge on a protein is based on the sum charge of its amino acids, and the pH of the buffer. Proteins are applied to a solid matrix such as an agarose gel, or a cellulose acetate membrane in a liquid buffer, and
484:
3). It is raised in the acute phase response. Depression of C3 occurs in autoimmune disorders as the complement system is activated and the C3 becomes bound to immune complexes and removed from serum. Fibrinogen, a beta-2 protein, is found in normal plasma but absent in normal serum. Occasionally,
552:
If the gamma zone shows an increase the first step in interpretation is to establish if the region is narrow or wide. A broad "swell-like" manner (wide) indicates polyclonal immunoglobulin production. If it is elevated in an asymmetric manner or with one or more peaks or narrow "spikes" it could
189:
In capillary electrophoresis, there is no solid matrix. Proteins are separated primarily by strong electroendosmotic forces. The sample is injected into a capillary with a negative surface charge. A high current is applied, and negatively charged proteins such as albumin try to move towards the
533:
are generally the only proteins present in the normal gamma region. Of note, any protein migrating in the gamma region will be stained and appear on the gel, which may include protein contaminants, artifacts, or certain medications. Depending on whether an agarose or capillary method is used,
556:
Polyclonal gammopathy is indicated by a "swell-like" elevation in the gamma zone, which typically indicates a non-neoplastic condition (although is not exclusive to non-neoplastic conditions). The most common causes of polyclonal hypergammaglobulinaemia detected by electrophoresis are severe
516:
samples will be seen in the beta gamma region. Fibrinogen, a beta-2 protein, is found in normal plasma but absent in normal serum. Occasionally, blood drawn from heparinized patients does not fully clot, resulting in a visible fibrinogen band between the beta and gamma globulins.
534:
interferences vary. Immunoglobulins consist of heavy chains (μ, δ, γ, α, and ε) and light chains (κ and λ). A normal gamma zone should appear as a smooth 'blush', or smear, with no asymmetry or sharp peaks. The gamma globulins may be elevated (
542:), or have an abnormal peak or peaks. Note that immunoglobulins may also be found in other zones; IgA typically migrates in the beta-gamma zone, and in particular, pathogenic immunoglobulins may migrate anywhere, including the alpha regions.
319:(AAT) constitutes most of the alpha-1 band. Alpha-1 antitrypsin has an SG group and thiol compounds may be bound to the protein altering their mobility. A decreased band is seen in the deficiency state. It is decreased in the
399:
Alpha-2 macroglobulin may be elevated in children and the elderly. This is seen as a sharp front to the alpha-2 band. AMG is markedly raised (10-fold increase or greater) in association with glomerular protein loss, as in
389:). Haptoglobin is raised as part of the acute phase response, resulting in a typical elevation in the alpha-2 zone during inflammation. A normal alpha-2 and an elevated alpha-1 zone is a typical pattern in hepatic
327:
from unregulated neutrophil elastase activity in the lung tissue. The alpha-1 fraction does not disappear in alpha 1-antitrypsin deficiency, however, because other proteins, including alpha-
549:. IgA deficiency occurs in 1:500 of the population, as is suggested by a pallor in the gamma zone. Of note, hypogammaglobulinema may be seen in the context of MGUS or multiple myeloma.
568:
A narrow spike is suggestive of a monoclonal gammopathy, also known as a restricted band, or "M-spike". To confirm that the restricted band is an immunoglobulin, follow up testing with
500:
has the most anodal mobility and typically migrates in the region between the beta and gamma zones also causing a beta/gamma fusion in patients with cirrhosis, respiratory infection,
396:
Haptoglobin/haemoglobin complexes migrate more cathodally than haptoglobin as seen in the alpha-2 – beta interzone. This is typically seen as a broadening of the alpha-2 zone.
132:(the fluid portion of the blood after the blood has clotted) is applied to either an acetate membrane soaked in a liquid buffer, or to a buffered agarose
604:
454:
101:
858:
Peralta, Ruben; Rubery, Brad A (July 30, 2012). Pinsky, Michael R; Sharma, Sat; Talavera, Francisco; Manning, Harold L; Rice, Timothy D (eds.).
545:
Hypogammaglobulinaemia is easily identifiable as a "slump" or decrease in the gamma zone. It is normal in infants. It is found in patients with
214:
is the major fraction in a normal SPEP. A fall of 30% is necessary before the decrease shows on electrophoresis. Usually a single band is seen.
932:"Serum alpha 2-macroglobulin and alpha 1-inhibitor 3 concentrations are increased in hypoalbuminemia by post-transcriptional mechanisms"
485:
blood drawn from heparinized patients does not fully clot, resulting in a visible fibrinogen band between the beta and gamma globulins.
104:(MGUS), or further investigate a discrepancy between a low albumin and a relatively high total protein. Unexplained bone pain, anemia,
431:
1166:
222:– two equally staining bands, the product of two genes. Some variants give rise to a wide band or two bands of unequal intensity but
1287:
823:
Hoang, Mai P; Baskin, Leland B; Wians, Frank H (1999). "Bisalbuminuria in an adult with bisalbuminemia and nephrotic syndrome".
600:
1062:
784:
753:
691:
1318:
1247:
1313:
1292:
1277:
658:
1396:
1282:
807:
176:
1323:
1272:
1212:
331:
and orosomucoid, also migrate there. As a positive acute phase reactant, AAT is increased in acute inflammation.
133:
766:
Medina-De la Garza CE, García-Hernández M, Castro-Corona MA. Visual mnemonics for serum protein electrophoresis
1227:
461:) comprises the beta-1. Increased beta-1 protein due to the increased level of free transferrin is typical of
1232:
1159:
1008:"Monitoring multiple myeloma patients treated with daratumumab: teasing out monoclonal antibody interference"
1129:
353:. These bands fuse and intensify in early inflammation due to an increase in alpha 1-antichymotrypsin, an
1207:
546:
1186:
1365:
1217:
350:
226:
of these variants is associated with disease. Increased anodic mobility results from the binding of
137:
1430:
1384:
1202:
1152:
346:
297:
270:
39:
144:
to separate the serum protein components into five major fractions by size and electrical charge:
96:. The most common indications for a serum protein electrophoresis test are to diagnose or monitor
70:
1138:
767:
539:
535:
462:
198:
109:
607:(MGUS), amyloidosis, plasma cell leukemia and solitary plasmacytomas also produce an M-spike.
1257:
1242:
573:
435:
366:
323:
and absence could indicate possible alpha 1-antitrypsin deficiency. This eventually leads to
285:
can lead to decrease in albumin level; due to its loss in the urine through a damaged leaky
1403:
1349:
505:
474:
439:
408:
239:
709:"Evaluation of a cellulose-acetate electrophoresis system for serum protein fractionation"
8:
1344:
1252:
984:
562:
354:
334:
316:
289:. An increase appears in severe alcoholics and in women during pregnancy and in puberty.
286:
190:
anode. Liquid buffer flows towards the cathode, and drags proteins with a weaker charge.
859:
572:, or immunodisplacement/immunosubtraction (capillary methods) is performed. Therapeutic
253:, is rare. A decreased level of albumin, however, is common in many diseases, including
1102:
1078:"Prevalence of Monoclonal Gammopathy of Undetermined Significance: A Systematic Review"
1077:
907:
882:
493:
481:
401:
374:
320:
282:
28:
836:
1425:
1222:
1107:
1039:
953:
948:
931:
912:
840:
803:
730:
687:
664:
654:
293:
983:
Tuazon, Sherilyn
Alvaran; Scarpaci, Anthony P (May 11, 2012). Staros, Eric B (ed.).
1267:
1097:
1089:
1029:
1019:
943:
902:
894:
832:
720:
646:
585:
149:
141:
97:
1237:
1175:
725:
708:
599:. Note that up to 8% of healthy geriatric patients may have a monoclonal spike.
526:
382:
153:
125:
45:
650:
569:
250:
219:
161:
89:
684:
Multiple
Myeloma and Related Serum Protein Disorders: An Electrophoretic Guide
1419:
1308:
1262:
589:
254:
157:
145:
113:
33:
Normal serum protein electrophoresis diagram with legend of different zones.
1111:
1043:
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1007:
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844:
668:
596:
513:
501:
420:
258:
243:
215:
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496:
is found in between the beta and gamma zones producing beta/gamma fusion.
55:
450:
378:
370:
328:
312:
308:
231:
129:
105:
898:
1034:
530:
509:
390:
386:
235:
617:
558:
466:
427:. There are low levels in inflammation and high levels in pregnancy.
324:
300:
may result in a sharp band between the albumin and the alpha-1 zone.
278:
274:
227:
93:
610:
Oligoclonal gammopathy is indicated by one or more discrete clones.
477:. Decreased beta-1 protein occurs in acute or chronic inflammation.
613:
470:
1144:
65:
577:
473:
therapy. Increased beta-1 protein due to LDL elevation occurs in
424:
211:
121:
645:. Methods in Molecular Medicine. Vol. 27. pp. 11–20.
621:
419:
Cold insoluble globulin forms a band here which is not seen in
772:
641:
Jenkins, Margaret A. (1999). "Serum
Protein Electrophoresis".
180:
Visual mnemonics for serum protein electrophoresis, right hand
261:, malabsorption, protein-losing nephropathy and enteropathy.
117:
116:
are also signs of multiple myeloma, and indications for SPE.
768:
https://www.tandfonline.com/doi/full/10.3402/meo.v18i0.22585
616:
may be seen as a band cathodal to gamma in myelomonocytic
553:
indicate clonal production of one or more immunoglobulins,
269:
Even staining in this zone is due to alpha-1 lipoprotein (
120:
must first be collected, usually into an airtight vial or
593:
581:
497:
458:
377:(haptoglobin is a suicide molecule which binds with free
561:, chronic liver disease, rheumatoid arthritis, systemic
964:
883:"An Electrophoretic Study of Nephrotic Sera and Urine"
783:
sfn error: no target: CITEREFHarris2012pages_117–123 (
273:– HDL). Decrease occurs in severe inflammation, acute
930:
Stevenson, FT; Greene, S; Kaysen, GA (January 1998).
929:
741:
1050:
778:
434:band in this zone. High levels are seen in type II
242:, and occasionally from tryptic digestion in acute
1134:Visual mnemonics for serum protein electrophoresis
880:
643:Clinical Applications of Capillary Electrophoresis
605:monoclonal gammopathy of undetermined significance
315:migrate together but orosomucoid stains poorly so
167:
1417:
1076:Wadhera, Rishi K.; Rajkumar, S. Vincent (2010).
822:
1075:
881:Longsworth, LG; Macinnes, DA (1 January 1940).
102:monoclonal gammopathy of uncertain significance
982:
407:AMG is mildly elevated early in the course of
340:
264:
88:) is a laboratory test that examines specific
1160:
857:
800:Protein Electrophoresis in Clinical Diagnosis
706:
682:Harris, Neil S.; Winter, William E. (2012).
681:
202:Serum protein electrophoresis normal results
184:
414:
385:and these complexes are rapidly removed by
1167:
1153:
634:
592:are not uncommon and usually give rise to
337:may bind to and retard the alpha-1 band.
193:
27:
1101:
1061:sfn error: no target: CITEREFHarris2012 (
1033:
1023:
947:
906:
752:sfn error: no target: CITEREFHarris2012 (
724:
345:Two faint bands may be seen representing
1288:Temperature gradient gel electrophoresis
1005:
588:spikes. chronic lymphatic leukaemia and
197:
175:
64:
640:
488:
128:is a laboratory technique in which the
1418:
1056:
747:
565:and other connective tissue diseases.
1148:
970:
797:
373:. There are typically low levels in
1391:
1319:Gel electrophoresis of nucleic acids
1248:Electrophoretic mobility shift assay
887:The Journal of Experimental Medicine
430:Beta lipoprotein forms an irregular
1314:DNA separation by silica adsorption
1293:Two-dimensional gel electrophoresis
1174:
13:
1278:Polyacrylamide gel electrophoresis
365:This zone consists principally of
14:
1442:
1122:
442:, and in the nephrotic syndrome.
1390:
1379:
1378:
1283:Pulsed-field gel electrophoresis
949:10.1046/j.1523-1755.1998.00734.x
802:. Hodder Arnold. pp. 1–14.
779:Harris, 2012 & pages 117–123
601:Waldenström's macroglobulinaemia
1324:Gel electrophoresis of proteins
1273:Moving-boundary electrophoresis
1213:Capillary electrochromatography
1069:
999:
985:"Serum protein electrophoresis"
976:
923:
874:
580:being the most common cause of
360:
303:
1228:Difference gel electrophoresis
851:
816:
791:
760:
700:
675:
423:because it is precipitated by
168:Acetate or gel electrophoresis
69:Schematic representation of a
1:
1329:Serum protein electrophoresis
1233:Discontinuous electrophoresis
837:10.1016/S0009-8981(99)00054-6
707:Kaplan, A; Savory, J (1965).
627:
620:in which it is released from
520:
249:Absence of albumin, known as
78:Serum protein electrophoresis
22:Serum protein electrophoresis
686:. Demos Medical. p. 5.
445:
136:matrix, or into liquid in a
7:
1208:Agarose gel electrophoresis
1128:Protein electrophoresis at
547:X-linked agammaglobulinemia
341:Alpha-1 – alpha-2 interzone
265:Albumin – alpha-1 interzone
10:
1447:
1187:History of electrophoresis
726:10.1093/clinchem/11.10.937
206:
1374:
1366:Electrophoresis (journal)
1358:
1337:
1301:
1218:Capillary electrophoresis
1195:
1182:
351:vitamin D binding protein
185:Capillary electrophoresis
52:
38:
26:
21:
1203:Affinity electrophoresis
798:Keren, David F. (2003).
651:10.1385/1-59259-689-4:11
415:Alpha-2 - beta interzone
347:alpha 1-antichymotrypsin
298:hepatocellular carcinoma
271:high density lipoprotein
218:individuals may produce
140:tube, and exposed to an
1082:Mayo Clinic Proceedings
194:Serum protein fractions
71:protein electrophoresis
1025:10.1515/cclm-2015-1031
540:hypogammaglobulinaemia
536:hypergammaglobulinemia
463:iron deficiency anemia
203:
181:
158:beta 1 and 2 globulins
110:chronic kidney disease
74:
1258:Immunoelectrophoresis
1243:Electrochromatography
1094:10.4065/mcp.2010.0337
1006:McCudden, C. (2016).
574:monoclonal antibodies
480:Beta-2 comprises C3 (
436:hypercholesterolaemia
367:alpha-2 macroglobulin
201:
179:
90:proteins in the blood
68:
1404:Analytical Chemistry
1350:Isoelectric focusing
936:Kidney International
825:Clinica Chimica Acta
506:rheumatoid arthritis
489:Beta-gamma interzone
475:hypercholesterolemia
440:hypertriglyceridemia
409:diabetic nephropathy
240:acetylsalicylic acid
1345:Electrical mobility
1253:Gel electrophoresis
899:10.1084/jem.71.1.77
563:lupus erythematosus
355:acute phase protein
335:Bence Jones protein
317:alpha 1 antitrypsin
292:The high levels of
713:Clinical Chemistry
494:C-reactive protein
482:complement protein
402:nephrotic syndrome
375:haemolytic anaemia
321:nephrotic syndrome
296:that may occur in
283:nephrotic syndrome
204:
182:
75:
1413:
1412:
1223:Dielectrophoresis
1012:Clin Chem Lab Med
973:, pp. 93–97.
860:"Hypoalbuminemia"
693:978-1-933864-75-4
508:(increased IgA).
369:(AMG or A2M) and
154:alpha-2 globulins
150:alpha-1 globulins
63:
62:
1438:
1394:
1393:
1382:
1381:
1268:Isotachophoresis
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1130:Lab Tests Online
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750:, pp. 9–16.
745:
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672:
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455:beta-lipoprotein
230:, nonesterified
142:electric current
98:multiple myeloma
56:edit on Wikidata
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1018:(6): 1095–104.
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393:and cirrhosis.
383:red blood cells
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162:gamma globulins
126:Electrophoresis
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1123:External links
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1088:(10): 933–42.
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998:
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719:(10): 937–42.
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570:immunofixation
538:), decreased (
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416:
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381:released from
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361:Alpha-2 zone
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259:malnutrition
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244:pancreatitis
223:
216:Heterozygous
210:
188:
171:
85:
81:
77:
76:
1057:Harris 2012
1035:2066/170995
748:Harris 2012
451:Transferrin
379:haemoglobin
371:haptoglobin
329:lipoprotein
313:antitrypsin
309:Orosomucoid
232:fatty acids
130:blood serum
106:proteinuria
1420:Categories
1196:Techniques
987:. Medscape
971:Keren 2003
862:. Medscape
628:References
531:antibodies
521:Gamma zone
510:Fibrinogen
391:metastasis
387:phagocytes
287:glomerulus
236:penicillin
991:2 October
866:2 October
618:leukaemia
559:infection
471:oestrogen
467:pregnancy
446:Beta zone
325:emphysema
279:cirrhosis
275:hepatitis
228:bilirubin
138:capillary
94:globulins
1426:Serology
1385:Category
1359:Journals
1112:20713974
1044:27028734
917:19870946
845:10437648
669:21374283
614:Lysozyme
432:crenated
281:. Also,
1397:Commons
1103:2947966
958:9453001
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735:4158264
624:cells.
603:(IgM),
578:Myeloma
425:heparin
212:Albumin
207:Albumin
122:syringe
92:called
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1338:Theory
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160:, and
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512:from
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118:Blood
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1063:help
1040:PMID
993:2013
954:PMID
913:PMID
868:2013
841:PMID
804:ISBN
785:help
754:help
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688:ISBN
665:PMID
655:ISBN
584:and
525:The
453:and
349:and
311:and
238:and
224:none
100:, a
82:SPEP
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1090:doi
1030:hdl
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903:PMC
895:doi
833:doi
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721:doi
647:doi
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