Atypical findings of protein studies using capillary zone electrophoresis.

We present two atypical cases of protein studies that were evaluated by immunofixation and immunosubtraction using capillary zone electrophoresis and high-resolution agarose gel electrophoresis. The first study showed an abnormal peak in the beta region by capillary zone electrophoresis that was located in the gamma region of the high-resolution agarose gel electrophoresis. Further investigation showed that this monoclonal protein was displaced due to binding with beta-lipoproteins. In the second case, a large peak was detected in the alpha-2 region and was shown by capillary zone electrophoresis to be a non-proteinaceous material that mimicked a paraprotein.     

血清蛋白电泳M蛋白阴性的骨髓瘤患者的特征学研究

目的：探讨血清蛋白电泳未检出单克隆条带,而免疫固定电泳可以检测出单克隆条带的多发性骨髓瘤患者的疾病的分型以及其生化指标的变化。方法：应用琼脂糖凝胶电泳方法和免疫沉淀方法对待测标本进行血清蛋白电泳和免疫固定电泳检测;应用全自动生化分析仪进行生化项目检测。结果：（1）实验组的骨髓瘤患者免疫固定电泳分型中,游离λ链显著高于对照组（P=0.04）;（2）实验组和对照组与健康对照组相比,α2/α1比值显著升高（P=0.046,P=0.00）;（3）实验组血清总蛋白和白蛋白水平显著低于对照组（P=0.00）,实验组血清钙离子水平显著高于对照组（P=0.012）。结论：2α/α1比值升高,总蛋白和白蛋白水平可能正常或下降,总血磷升高的怀疑为多发性骨髓瘤的患者,应推荐做免疫固定电泳检测。     

Identification and quantification of a monoclonal IgM euglobulin: pitfalls and methodologic instruction

In a patient with Waldenstr?m's macroglobulinemia routine agarose gel electrophoresis and immunofixation disclosed an IgM-kappa M-component with beta-mobility, but failed to unveil a major monoclonal IgM-kappa euglobulin. This, however, was demonstrated when the procedures were performed in a buffer with NaCl-ion-strength of 0.1 M. Similarly, IgM quantification by routine nephelometry and electroimmunoassay led to dramatically inconsistent results. By a simple agarose gel electrophoresis in NaCl 0.1 M the IgM euglobulin was quantified proportional to the serum concentration of albumin. The identification and classification of an M-component, being exceedingly important diagnostically, may be missed, when solely routine electrophoresis and immunofixation is employed. As regards the quantification of M-components, these are not merely pathologic immunoglobulins nosologically. Also, biochemically they differ from normal, polyclonal immunoglobulins, resulting in immunological techniques being inapplicable for their quantification. For quantitative purposes, a simple, comparative agarose gel electrophoresis is described.     

免疫固定电泳在多发性骨髓瘤辅助诊断中的临床价值

目的 探讨血清免疫固定电泳在多发性骨髓瘤(MM)的辅助诊断与治疗中的临床价值.方法 以2017年1月至2019年10月就诊于新疆维吾尔自治区人民医院的74例MM患者作为研究对象,分别取血清进行免疫固定电泳与蛋白电泳,收集两种电泳方法的M蛋白检测结果 并进行统计学处理.结果 74例MM患者中,血清蛋白电泳检出M蛋白51例...     

血清免疫固定电泳、蛋白电泳、免疫球蛋白及轻链定量对多发性骨髓瘤临床诊断价值探讨

目的探讨血清蛋白电泳、总蛋白、免疫球蛋白及轻链、蛋白电泳、免疫固定电泳测定对多发性骨髓瘤(MM)的诊断价值。方法对89例多发性骨髓瘤患者的血清总蛋白、免疫球蛋白及轻链、蛋白电泳、免疫固定电泳和尿轻链κ、λ的检测结果进行统计分析。结果 89例MM患者38例患者血清总蛋白含量升高;55例免疫球蛋白及轻链定量显著升高;血清蛋白电泳56例检出M带,而免疫固定电泳中89例均为阳性结果。结论免疫固定电泳检是MM诊断的敏感方法 ,应作为MM的常规检测方法 。     

Inheritance of human α1-acid glycoprotein (orosomucoid) variants

Although variants of sialic acid-free alpha(1)-acid glycoprotein have been described in human beings, the mode of inheritance of these types has not been reported previously. With the use of a new technique of immunofixation after agarose gel electrophoresis of neuraminidase-treated whole serum, the present study demonstrates that the types of alpha(1)-acid glycoprotein variants in family members are consistent with inheritance as autosomal traits with codominant expression. Gene frequencies have been determined for several ethnic groups. Of a total of 11 maternal-cord serum pairs, seven were discordant types, indicating that the fetus synthesizes alpha(1)-acid glycoprotein and confirming a previous report that there is no significant transplacental passage of this protein.     

多发性骨髓瘤血清蛋白电泳与免疫学分型

目的对38例多发性骨髓瘤(MM)患者血清蛋白电泳、免疫电泳及免疫球蛋白进行定量分析,以探讨M蛋白、MM型别分布及其在MM中的临床诊断价值。方法血清蛋白和免疫电泳采用全自动琼脂糖凝胶电泳仪进行电泳和扫描;免疫球蛋白定量采用Beckman CS-免疫化学分析仪,以全自动方式进行速率散射比浊分析,测各样本的免疫球蛋白(IgG、IgA、Ig M)含量。结果38例MM全部检出M带,22例MM尿液检出本-周氏蛋白,检出阳性率为57.90%。血清免疫电泳分型IgGκ型14例,占36.84%;IgGλ型10例,占26.32%;IgAκ型5例,占13.16%;IgAλ型6例,占15.79%;Ig Mλ型3例,占7.89%,为最少。结论血清蛋白及免疫电泳能快速、准确地进行MM分型,对MM的诊断、病情判断、治疗及预后评估均具有重要的临床价值。     

Rapid detection and initial characterization of genetic variants of human serum albumin.

We have studied the detection and classification of genetic variants of human serum albumin by electrophoresis. Samples from 10 patients who were heterozygous for eight different albumin variants were studied by two methods. In agarose gel electrophoresis, each of these variants has an abnormal mobility and can be classified on the basis that structural changes at the N-terminus abolish 63Ni binding. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole serum, glycosylated variants are easily detected because of their greater apparent molecular mass.     

31例轻链型多发性骨髓瘤实验室指标分析

目的探讨实验室检测指标在轻链型多发性骨髓瘤(MM)诊治中的应用价值。方法收集轻链型MM患者31例及对照组60例,对照组为健康者20例(健康组),以及该院诊断的IgGκ型和IgAκ型MM患者各20例。检测免疫球蛋白、血清蛋白电泳、免疫固定电泳、尿素氮(UREA)、肌酐(Cr)、β_2微球蛋白,并进行流式细胞免疫表型检测及骨髓细胞学检查。结果血清蛋白电泳检测到M蛋白的阳性率为29.03%,Durie-Salmon临床分期Ⅱ期与Ⅲ期M蛋白水平差异有统计学意义(P0.05);免疫固定电泳结果均需用琼脂糖凝胶电泳法加做IgD及IgE抗体复查;轻链型MM患者与健康者的IgG、IgA、IgM、UREA、Cr、β_2微球蛋白比较,差异有统计学意义(P0.05);MM细胞形态特征改变显著,细胞抗原表达以CD38、CD138、CD56为主。结论免疫球蛋白定量及免疫固定电泳可作为M蛋白的筛查方法,血清蛋白电泳可用于患者的临床分期和疗效观察;骨髓细胞学检查对轻链型MM起到重要的诊断作用,轻链型MM比其他类型MM有更加严重的肾功能损伤。     

应用高效毛细管电泳筛检和鉴定血清中M蛋白

目的:应用毛细管区带电泳法(CZE)和毛细管区带电泳-免疫削减法(IS-CZE)对M蛋白进行筛检和鉴定。方法:运用CZE和琼脂糖凝胶电泳(AGE)分别对532例血清进行M蛋白的筛检;用IS-CZE和免疫固定电泳(IFE)对M蛋白阳性的64例标本进行免疫分型。结果:与AGE相比较,CZE筛检M蛋白的阳性率为96.9%;与IFE相比较,IS-CZE分型的符合率为89.1%;自包被固相抗体能满足试验要求。结论:毛细管区带电泳可高效、快速完成M蛋白的筛检和鉴定,适合临床推广。     

Use of immunoglobulin heavy- and light-chain measurements compared with existing techniques as a means of typing monoclonal immunoglobulins

Using automated measurements of kappa and lambda light chains and IgG, IgA, and IgM, we assessed the utility of the kappa/lambda ratio and the heavy chain/light chain ratio in characterizing monoclonal immunoglobulins previously identified by sensitive electrophoresis on agarose gel and typed by immunofixation or immunoelectrophoresis. We examined 348 selected samples, of which 165 contained monoclonal components, finding that 93.4% were detected and 89% correctly typed by this approach. Eight samples were shown to contain monoclonal light chains that were not visible as bands on electrophoresis but demonstrated abnormalities of the kappa/lambda or heavy chain/light chain ratio.     

免疫固定电泳在多发性骨髓瘤中的应用

目的探讨免疫固定电泳技术（IFE）在鉴定多发性骨髓瘤诊断中的应用。方法采用免疫固定电泳技术检测50例多发性骨髓瘤患者血清标本。结果其中45例多发性骨髓瘤检出M蛋白,M蛋白分型为IgGK型16例,IgGh型13例,IgAK型6例,IgAh型10例,单纯入型2例;3例未检出M带。结论采用免疫固定电泳技术诊断多发性骨髓瘤,沉淀带易识别,操作简单、快速。对鉴定各类型M蛋白有重要价值。     

Macro creatine kinase in a case of carnitine palmitoyltransferase deficiency

A stout man was admitted to the hospital with acute rhabdomyolysis associated with macro creatine kinase (macro-CK, EC 2.7.3.2). This anomaly of CK was detected by gel electrophoresis as an atypical band between CK-MB and CK-MM, classified according to Stein's criteria (Clin Chem 1982; 28:19-24) as type 1, and identified by immunofixation electrophoresis as containing CK isoenzymes MM and MB and immunoglobulin A. Muscle biopsy showed that the etiology of rhabdomyolysis in this case was deficiency of carnitine palmitoyltransferase (CPT, EC 2.3.1.21) in the muscle. We report the first observation of macro-CK in a case of CPT deficiency; its presence may result from recurrent rhabdomyolytic attacks owing to CPT deficiency, and may suggest underlying enzymic abnormality in muscle.     

Bonded-phase extraction of plasma tricyclic antidepressant drugs for gas chromatographic analysis

Demonstration of an oligoclonal immunoglobulin pattern in cerebrospinal fluid by a commercial agarose gel electrophoresis system and immunofixation were evaluated in the service clinical laboratory of a university hospital. In 303 patients, 45 with clinically definite multiple sclerosis and 209 with other neurologic diseases, the sensitivity of oligoclonal banding for multiple sclerosis was 71%, and the specificity, 83%. Oligoclonal banding was frequent in inflammatory disease, tumor/pseudotumor or vascular diseases of the central nervous system (35%, 36% and 26%, respectively) and less frequent in degenerative central nervous system disease and peripheral neuropathy (5.2 % and 15%). No patient with non-neurologic disease had oligoclonal banding. The addition of an immunochemical step (immunofixation) did not increase sensitivity and only minimally increased specificity. It did permit distinction in selected cases between immunoglobulin and other molecules with identical electrophoretic mobility.     

Alpha 1-antitrypsin: the effect of anticoagulants on the trypsin inhibitory capacity, concentration and phenotype

The effects of anticoagulants on the determination of both trypsin inhibitory capacity and the concentration of alpha 1-antitrypsin measured by radial immunodiffusion, and on the alpha 1-antitrypsin phenotype were investigated. These results were compared with those obtained for serum. The following anticoagulants were investigated: sodium citrate; sodium oxalate; buffered citrate; potassium oxalate/sodium fluoride; sodium heparin; and potassium EDTA. It was found that plasmas from all of the anticoagulants, except sodium heparin, resulted in apparently significant decreases of both trypsin inhibitory capacity and concentration of alpha 1-antitrypsin measured by radial immunodiffusion, relative to serum. These decreases were not simply due to dilution by anticoagulants. Using both acid starch gel electrophoresis followed by immunofixation and isoelectric focusing in agarose, no interference was found in the phenotype determination. It is concluded that serum should be used to measure the trypsin inhibitory capacity or the concentration of alpha 1-antitrypsin by radial immunodiffusion, although plasma is also suitable provided that sodium heparin is used as the anticoagulant. The alpha 1-antitrypsin phenotype can be determined with either serum or any of the plasma. None of the anticoagulants employed in this study was present in excess.     

Detection of oligoclonal immunoglobulins in cerebrospinal fluid by an immunofixation-peroxidase method.

BACKGROUND: The detection of intrathecal synthesis of immunoglobulins is used in the diagnosis of multiple sclerosis (MS). We tested the semiautomated immunofixation-peroxidase (IFPOD) technique, which uses high-resolution agarose gel electrophoresis (HRAGE) directly followed by immunofixation with a peroxidase-labeled anti-IgG antiserum to detect oligoclonal immunoglobulins in cerebrospinal fluid (CSF). METHODS: We analyzed 230 consecutive matched serum/CSF pairs that arrived in the laboratory over a 6-month period with both IFPOD and our routine techniques, immunofixation electrophoresis (IFE) and HRAGE. For IFPOD, CSF samples were not concentrated before testing. RESULTS: Among the 230 samples were 12 clinically definite MS, 33 clinically probable, and 20 clinically possible MS samples. IFPOD and HRAGE + IFE each detected oligoclonal IgG in CSF in 10, 16, and 7 cases of these respective groups. For clinically definite MS, sensitivity and specificity (95% confidence intervals) were, respectively, 83% (51-97%) and 79% (73-84%). CONCLUSIONS: The IFPOD technique performs comparably to other analytical methods, without the requirement for sample concentration, and may represent an attractive alternative in testing for intrathecal immunoglobulin synthesis.     

28例多发性骨髓瘤的血清免疫学分析

目的　探讨多发性骨髓瘤(multiplemyeloma,MM)的血清免疫学特征。　方法　对28例MM患者的血清进行琼脂糖凝胶电泳及电泳扫描,血清蛋白和免疫球蛋白定量,以及免疫固定电泳。　结果　27例(96.4%)MM患者血清蛋白电泳可显示M带,IgG型、IgM型多在γ区,IgA型多在β区。不同型别MM免疫固定电泳图谱各异,IgG、IgA、IgM和单纯游离轻链的比例分别为46.4%、21.4%、17.9%、10.7%。在轻链型别中,IgG、IgM类M蛋白以κ型为主,单纯轻链型以λ型为主。免疫球蛋白定量显示M蛋白所属免疫球蛋白部分显著增高,非M蛋白所属免疫球蛋白显著降低。　结论　通过上述四种免疫化学综合实验对MM进行免疫分型,可对MM作出准确的实验室诊断。     

Restricted electrophoretic heterogeneity of immunoglobulin light chains in urine: a cause for confusion with Bence Jones protein

The detection of Bence Jones protein, an important part of the investigation of suspected myeloma, is most commonly done by agarose or cellulose nitrate electrophoresis followed by immunofixation. Bence Jones protein is recognized as single or multiple bands of one type of light chain. Unfortunately, improvements in sensitivity of these techniques (use of high-affinity antisera and higher resolution electrophoresis) frequently allow detection of multiple light chain bands in the urine of patients who do not have a B-cell dyscrasia. The bands are usually kappa, although they may be accompanied by lambda bands. This pattern may lead to the misdiagnosis of Bence Jones protein and oligoclonal light chain production in patients. Here we show that this pattern is produced by polyclonal light chains; it is present in the urine of all patients with a tubular proteinuria of any etiology and may be induced in healthy individuals by blocking their renal tubular protein reabsorption. Polyclonal light chains separate into monomers and dimers on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and into four major bands with many minor bands by isoelectric focusing. This difference in charge and possibly size results in the banding pattern seen on good-quality electrophoresis and immunofixation.     

Immunofixation. III. Application to the study of monoclonal proteins.

We have shown how immunofixation can be successfully used to study proteins exhibiting electrophoretic polymorphism. An analogous situation is seen in the disorders of the immune system that result in the production of variable quantitites of homogeneous immunoglobulins. Immunoelectrophroesis has been heavily used--sometimes unsuccessfully because of poor resolution--to study these materials. The appearance of a monoclonal immunoglobulin band is identical in both immunofixation and standard gel electrophoresis, a feature that allows unambiguous identification of a protein as to its heavy-chain class and light-chain type. Because antigen and antibody are placed in contact almost immediately after electrophoresis, interfering diffusion does not occur and even extremely small bands can be demonstrated and characterized.     

Detection of beta 2-transferrin with agarose gel electrophoresis, immunofixation and silver staining in cerebrospinal fluid, secretions and other body fluids

A method is described for the detection of beta 2-transferrin (asialotransferrin) in 1 microliter cerebrospinal fluid. Owing to its low content of sialic acid, beta 2-transferrin migrates more slowly than the main transferrin component (beta 1-transferrin) in electrophoresis. The high sensitivity and specificity of the test depend on immunofixation of the electrophoretically separated transferrin in the agarose gel, and on the visualization of the immune complex by staining with alkaline silver nitrate solution. Since beta 2-transferrin occurs practically only in cerebrospinal fluid and not in other body fluids, its detection in secretions from the nose or ear can be used for the diagnosis of rhinoor otoliquorrhoea. Even under unfavourable conditions, 10% cerebrospinal fluid can be detected in a secretion. Operator time for investigation of a secretion sample is 110 minutes and the analysis time is 5 hours. The direct costs are 95 German marks. Forty two secretions have been investigated so far. Eighteen were shown to contain cerebrospinal fluid, and this was confirmed by subsequent operative findings on the patients.