Free accessReview articleFirst published online 1985-7
High Electroendosmotic Agarose Electrophoresis and Nitrocellulose Immobilisation for the Detection of Oligoclonal Bands in Unconcentrated Cerebrospinal Fluid
A sensitive (1 μL of unconcentrated sample), specific (IgG Fc) and economical (10 p/test) method for the detection of oligoclonal bands in cerebrospinal fluid is presented. Using commonly available technology (agarose gel electrophoresis) and giving results within 6 h, the approach used here is a powerful tool in the investigation of immunoglobulin abnormalities in many biological fluids.
References
1.
KabatEAMooreDHLandhowH.An electrophoretic study of the protein components in cerebrospinal fluid and their relationship to serum proteins. J Clin Invest1942; 21: 571–7.
2.
LowenthalAVansandeMKarcherD.The differential diagnosis of neurological disease by fractionating electrophoretically the cerebrospinal fluid proteins. J Neurochem1960; 6: 51–60.
3.
KarcherAVansandeMLowenthalA.Microelectrophoresis in agar gel of the proteins of the cerebrospinal fluid and central nervous system. J Neurochem1959; 4: 135.
4.
LinkHMullerR.Immunoglobulins in multiple sclerosis and infections of the nervous system. Arch Neurol1971; 25: 326–44.
5.
MattsonDHRoosRPArnasonBGW.Comparison of agar gel electrophoresis and isoelectric focussing in multiple sclerosis and subacute sclerosing panencephalitis. Ann Neurol1981; 9: 34–41.
6.
VandvikBSkredeS.Electrophoretic examination of cerebrospinal fluid proteins in multiple sclerosis and other neurological diseases. Europ Neurol1973; 9: 224–41.
7.
JohnsonKPArrigoSCNelsonBJAgarose electrophoresis of cerebrospinal fluid in multiple sclerosis. Neurology1977; 27: 273–7.
8.
GersonBKrolokowskiFJGersonIM.Two agarose electrophoresis systems for demonstration of oligoclonal bands in cerebrospinal fluid compared. Clin Chem1980; 26: 343–4.
9.
PatyDWDonnellyMBernardoME.CSF electrophoresis, an adaptation using cellulose acetate for the identification of oligoclonal banding. Can J Neurol Sci.1978; 5: 297–9.
10.
ThompsonEJKaufmanPShortmanRCOligoclonal immunoglobulins and plasma cells in spinal fluid of patients with multiple sclerosis. Br Med J1979; i: 16–17.
11.
GauthierFChasseJFMourayH.A simple characterisation of oligoclonal immunoglobulins in unconcentrated cerebrospinal fluid using polyacrylamide gel electrophoresis. Clin Chim Acta1983; 127: 407–11.
12.
ThompsonEJJohnsonMH.Electrophoresis of CSF proteins. Br J Hosp Med.1982; 26: 600–8.
13.
DelmotteP.Comparative results of agar gel electrophoresis and isoelectric focussing examination of the gamma globulins of the cerebrospinal fluid. Acta Neurol Belg1972; 72: 226–34.
14.
KjellinKGVesterbergO.Isoelectric focussing of CSF proteins in neurological diseases. J Neurol Sci.1974; 23: 199–213.
15.
KleineTOStrohJ.Neue microelectrophoreses für nativen lumballiquor unterschede in pherogramm von nativen and konzentrierten proteinen. Z Clin Chem Klin Biochem1974; 12: 73–80.
16.
LindstedtGLundbergP-A.Loss of tubular proteinuria pattern during concentrating with a commercial membrane filter cell (Minicon B-15 system). Clin Chim Acta1974; 56: 125–6.
17.
KerenyiLGallyasF.A highly sensitive method for demonstrating proteins in electrophoretic, immunoelectrophoretic and immunodiffusion studies. Clin Chim Acta1972; 38: 564–7.
18.
KarcherDLowenthalAVan SoomG.Cerebrospinal fluid protein electrophoresis without prior concentration. Acta Neurol Belg1979; 79: 335–7.
19.
MerrillCRGoldmanDSedmanSAUltrasensitive stain for proteins in polyacrylamide gels shows regional variations in CSF proteins. Science1981; 211: 1437–8.
20.
WilloughbyEWLambertA.A Sensitive silver stain for proteins in agarose gels. Annal Biochem1983; 130: 353–8.
21.
CawleyLPMinardBJTourtellotteeWWImmunofixation electrophoretic techniques applied to identification of proteins in serum and cerebrospinal fluid. Clin Chem1976; 22: 1262–8.
22.
MattsonDHRoosRPArnasonBGW.Immunoperoxidase staining of cerebrospinal fluid IgG in isoelectric focussed gels—a sensitive new technique. J Neurosci Meth1980; 3: 67–75.
23.
ChazotGLasneYBenzeraraORadio-immunofixation une novelle technique de characterisation des immunogulines dans de liquide cepahlo-rachidien non concentre. Rev Neurol (Paris)1980; 136: 783–6.
24.
LasneBBenzeraraIChazotGCreysselR.A sensitive method for characterisation of oligoclonal immunoglobulins in unconcentrated cerebrospinal fluid. J Neurochem1981; 36: 1872–4.
25.
KostulosKLinkH.Agarose isoelectric focussing of unconcentrated CSF and radio-immunofixation for the detection of oligoclonal bands in patients with multiple sclerosis and other neurological diseases. J Neurol Sci.1982; 54: 117–27.
26.
JohnsonMHWalkerRWHKeirGIdentification of protein bands in polyacrylamide gel by protein blotting. Biochim Biophys Acta1982; 718: 121–4.
27.
WalkerRWHKeirGJohnsonMHA rapid method for detecting oligoclonal IgG in unconcentrated CSF, by agarose isoelectric focussing, transfer to cellulose nitrate and immunoperoxidase staining. J Neuroimmunol1983; 4: 141–8.
28.
JeppssonJ-OLaurellC-BFranzenB.Agarose gel electrophoresis. Clin Chem1979; 25: 629–38.
29.
LisakRPKeshgegianAA.Oligoclonal immunoglobulins in cerebrospinal fluid in multiple sclerosis. Clin Chem1980; 111: 1340–1.
30.
PoloniMRocchelliBScelsiRIntrathecal IgG synthesis in multiple sclerosis and other neurological diseases: a comparative evaluation by IgG-index and isoelectric focussing. J Neurol1979; 221: 245–55.
31.
GersonBCohenSRGersonIMMyelin basic protein, oligoclonal bands and IgG in cerebrospinal fluid as indicators of multiple sclerosis. Clin Chem1981; 27: 1974–7.
32.
LaurellC-B.Composition and variation of the gel electrophoretic fractions of plasma, cerebrospinal fluid and urine. Scand J Clin Lab Invest1972; Suppl. 12429: 71.
33.
LinkH.Immunoglobulin G and low molecular weight proteins in human cerebrospinal fluid. Acta Neurol Scand1967; 43: Suppl. 28: 1–136.
34.
HochwaldGMThorbeckGJ.Trace proteins in cerebrospinal fluid and other biological fluids. II. Effect of storage and enzymes on the electrophoretic mobility of β-trace and γ-trace proteins in cerebrospinal fluid. Clin Chim Acta1963; 8: 678–84.
35.
LaurenziMA.Immunochemical characterisation of immunoglobulins and viral antibodies synthesised within the CNS in patients with multiple sclerosis and controls. Acta Neurol Scand1981; 63: Suppl. 84: 1–84.
36.
FossardCDaleGLatnerAL.Separation of the proteins of cerebrospinal fluid using gel electrofocussing followed by electrophoresis. J Clin Path1970; 23: 586–9.
37.
DelmotteP.Gel isoelectric focussing of cerebrospinal fluid proteins: a potential diagnostic tool. Z Klin Chem Klin Biochem1971; 9: 334–6.
38.
LaurenziMALinkH.Comparison between agarose gel electrophoresis and isoelectric focussing of CSF for demonstration of oligoclonal immunoglobulin bands in neurological disorders. Acta Neurol Scand1978; 58: 148–56.
39.
OlssonJ-ENilssonK.Gamma globulin of cerebrospinal fluid and serum in multiple sclerosis: isoelectric focussing on polyacrylamide gel and agar electrophoresis. Neurology1979; 29: 1383–91.
40.
HersheyLATrotterJL.The use and abuse of cerebrospinal fluid IgG profile in the adult: a practical evaluation. Ann Neurol1980; 8: 426–34.
41.
RosenAEkKAmanP.Agarose isoelectric focussing of native human immunoglobulin M and α2-macroglobulin. J Immunol Meth1979; 28: 1–11.
42.
SavarisCAZamchekN.Isoelectric focussing in agarose. J Immunol Meth1979; 29: 91–6.
43.
LinkHKostulasV.Utility of isoelectric focussing of cerebrospinal fluid and serum on agarose evaluated for neurological patients. Clin Chem1983; 29: 810–15.
44.
AmblerJWalkerG.Isoelectric focussing of serum proteins on modified thin cellulose acetate membranes. Clin Chem1979; 25: 1320–2.
45.
GianazziERighettiPG.Facts and artefacts in isoelectric focussing. In: RighettiPGAllenRC, eds. Electrophoresis '79. Berlin: de Gruyter, 1980; 129–40.
46.
FelgenhauerKMohrmannH.Evaluation of immunoglobulin diversity. In: AllenRCArnaudP, eds. Electrophoresis '81. Berlin: de Gruyter, 1981; 427–32.
47.
ChrambachAder LanB AnMohrmanHToward an improved immunoglobulin analysis by gel electrophoresis and electrofocussing. Electrophoresis1981; 2: 279–87.
48.
WalkerRWHKeirGThompsonEJ.Assessment of cerebrospinal fluid immunoglobulin G patterns after isoelectric focussing: use of kappa and lambda light chain immunoperoxidase staining. J Neurol Sci.1983; 58: 123–34.
49.
NilssonKOlssonJ-E.Analysis for cerebrospinal fluid proteins by isoelectric focussing on polycrylamide gel. Clin Chem1978; 24: 1134–9.
50.
LaurenziMAMavraMKam-HansenSOligoclonal IgG and free light chains in multiple sclerosis demonstrated by thin-layer polyacrylamide gel isoelectric focussing and immunofixation. Ann Neurol1980; 8: 241–7.
51.
HoseinZZJohnsonKP.Isoelectric focussing of cerebrospinal fluid proteins in the diagnosis of mutiple sclerosis. Neurol (Minneap)1981; 31: 70–6.
52.
ZettervalOLinkH.Electrophoretic distribution of kappa and lambda immunoglobulin light chain determinants in serum and cerebrospinal fluid in multiple sclerosis. Clin Exp Immunol1970; 7: 365–72.
53.
MattsonDHRoosRHopperJELight chain composition of CSF oligoclonal IgG bands in multiple sclerosis and subacute sclerosing panencephalitis. J Neuroimmunol1982; 3: 63–76.
54.
GoodlandFCThompsonEJ.A comparison on cellulose acetate immunofixation with polyacrylamide gel electrophoresis for the detection of oligoclonal bands in unconcentrated cerebrospinal fluid. J Clin Path1983; 36: 1309–11.