scholarly journals Computer-supported Detection of M-Components and Evaluation of Immunoglobulins after Capillary Electrophoresis

2001 ◽  
Vol 47 (1) ◽  
pp. 110-117 ◽  
Author(s):  
Magnus Jonsson ◽  
Joyce Carlson ◽  
Jan-Olof Jeppsson ◽  
Per Simonsson
Keyword(s):  
Capillary Electrophoresis ◽  
Decision Support ◽  
Protein Separation ◽  
Serum Proteins ◽  
Cost Effective ◽  
Clinical Samples ◽  
Serum Samples ◽  
Computerized Decision Support ◽  
Cost Effective Method ◽  
Mathematical Algorithms

Abstract Background: Electrophoresis of serum samples allows detection of monoclonal gammopathies indicative of multiple myeloma, Waldenström macroglobulinemia, monoclonal gammopathy of undetermined significance, and amyloidosis. Present methods of high-resolution agarose gel electrophoresis (HRAGE) and immunofixation electrophoresis (IFE) are manual and labor-intensive. Capillary zone electrophoresis (CZE) allows rapid automated protein separation and produces digital absorbance data, appropriate as input for a computerized decision support system. Methods: Using the Beckman Paragon CZE 2000 instrument, we analyzed 711 routine clinical samples, including 95 monoclonal components (MCs) and 9 cases of Bence Jones myeloma, in both the CZE and HRAGE systems. Mathematical algorithms developed for the detection of monoclonal immunoglobulins (MCs) in the γ- and β-regions of the electropherogram were tested on the entire material. Additional algorithms evaluating oligoclonality and polyclonal concentrations of immunoglobulins were also tested. Results: CZE electropherograms corresponded well with HRAGE. Only one IgG MC of 1 g/L, visible on HRAGE, was not visible after CZE. Algorithms detected 94 of 95 MCs (98.9%) and 100% of those visible after CZE. Of 607 samples lacking an MC on HRAGE, only 3 were identified by the algorithms (specificity, 99%). Algorithms evaluating total gammaglobulinemia and oligoclonality also identified several cases of Bence Jones myeloma. Conclusions: The use of capillary electrophoresis provides a modern, rapid, and cost-effective method of analyzing serum proteins. The additional option of computerized decision support, which provides rapid and standardized interpretations, should increase the clinical availability and usefulness of protein analyses in the future.

scholarly journals Quantification of Serum Proteins Using Capillary Electrophoresis

1995 ◽  
Vol 32 (5) ◽  
pp. 493-497 ◽  
Author(s):  
M A Jenkins ◽  
M D Guerin
Keyword(s):  
Capillary Electrophoresis ◽  
Gel Electrophoresis ◽  
Serum Proteins ◽  
Clinical Laboratory ◽  
Charged Particles ◽  
Protein Electrophoresis ◽  
Agarose Gel Electrophoresis ◽  
Serum Samples ◽  
Routine Clinical Laboratory ◽  
Sample Dilution

Capillary electrophoresis is a technique that can be automated for the separation of charged particles. By investigating suitable sample dilution and injection time and adhering to a strict washing procedure we have been able to quantify paraproteins in serum samples. This has enabled us to use the technique of capillary electrophoresis for the provision of serum protein electrophoresis in a routine clinical laboratory. We present our findings of 260 serum samples, which included 76 samples with paraproteins analysed by both capillary electrophoresis (EC) and high resolution agarose gel electrophoresis (HRAGE). CE was able to detect all the monoclonal bands detected by HRAGE, and, in particular, better able to detect IgA monoclonal bands occurring in the beta region. The major advantages of CE over HRAGE relate to the automated nature of CE with the elimination of the need for a densitometer.

scholarly journals High-Quality, Cost-Effective Strategy for Detection of Autoantibodies to Extractable Nuclear Antigens

2001 ◽  
Vol 8 (3) ◽  
pp. 471-474 ◽  
Author(s):  
Tri G. Phan ◽  
Watson W. S. Ng ◽  
Dana Bird ◽  
Kara Smithers ◽  
Vicky Wong ◽  
...  
Keyword(s):  
Cost Effective ◽  
Turnaround Time ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Quality Cost ◽  
Nuclear Antigens ◽  
Cost Effective Method ◽  
Meaningful Result ◽  
Clinical Diagnoses

ABSTRACT We evaluated methods for the detection of autoantibodies to extractable nuclear antigens (ENAs) to determine the strategy that yielded the most cost effective and clinically meaningful result. We prospectively compared counterimmunoelectrophoresis (CIEP) with and without serum prediffusion (SPD) and found that SPD significantly improved the quality of precipitation lines. This resulted in a decreased requirement for repeat testing and, consequently, was associated with a significant decrease in reagent costs and specimen turnaround time. We also retrospectively compared reactivity by CIEP, CIEP plus SPD, enzyme-linked immunosorbent assay (ELISA), and line immunoassay (LIA) of 52 serum samples that were previously determined to be positive for ENAs, and we correlated the results with clinical diagnoses. There was significant agreement among CIEP, CIEP plus SPD, ELISA, and LIA for the detection of anti-SS-A, anti-SS-B and anti-RNP. In general, CIEP, CIEP plus SPD, and LIA correlated better with the clinical diagnoses than ELISA, even though ELISA detected anti-ENAs more often than the other methods. CIEP plus SPD is therefore the most cost effective method for the identification of clinically meaningful ENAs. Based on our experience, we now screen for ENAs by CIEP, and positive samples are then typed by CIEP plus SPD. Samples that are difficult to interpret are then further assessed by an alternative method.

Quantitative analysis of serum proteins separated by capillary electrophoresis

1993 ◽  
Vol 39 (4) ◽  
pp. 689-692 ◽  
Author(s):  
J W Kim ◽  
J H Park ◽  
J W Park ◽  
H J Doh ◽  
G S Heo ◽  
...  
Keyword(s):  
Capillary Electrophoresis ◽  
Gel Electrophoresis ◽  
Serum Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Serum Samples ◽  
Polyclonal Gammopathy ◽  
Quantitative Analyses

Abstract The possibility of open tubular capillary electrophoresis for clinical diagnostic use is examined. Capillary electrophoresis was performed in an untreated 50 microns (i.d.) x 100 cm (65 cm to detector) capillary with detection of absorbance at 200 nm. Conditions for the separation of serum proteins without adsorption to the capillary surface were established. Quantitative analyses of serum samples from 38 patients with liver cirrhosis, nephrotic syndrome, or polyclonal gammopathy by capillary electrophoresis were done and the results were compared with those by conventional agarose gel electrophoresis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. All samples were analyzed in duplicate. We evaluated linearity of response, within-run CV, and the correlation between capillary electrophoresis and agarose gel electrophoresis.

A Rapid and Cost-Effective Method for Genotyping Genome-Edited Animals: A Heteroduplex Mobility Assay Using Microfluidic Capillary Electrophoresis

2016 ◽  
Vol 43 (5) ◽  
pp. 341-348 ◽  
Author(s):  
Vanessa Chenouard ◽  
Lucas Brusselle ◽  
Jean-Marie Heslan ◽  
Séverine Remy ◽  
Séverine Ménoret ◽  
...  
Keyword(s):  
Capillary Electrophoresis ◽  
Cost Effective ◽  
Heteroduplex Mobility Assay ◽  
Cost Effective Method

scholarly journals Dextranol: A better lyoprotectant

2018 ◽  
Author(s):  
Bryan Joshua Jones ◽  
Advitiya Mahajan ◽  
Alptekin Aksan
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Room Temperature ◽  
Elevated Temperatures ◽  
Serum Proteins ◽  
Specific Antigen ◽  
Clinical Samples ◽  
Serum Samples ◽  
Protein Molecules ◽  
Anomeric Carbon

Dextranol, a reduced dextran, prevents damage to stored dry protein samples that unmodified dextran would otherwise cause. Lyoprotectants like the polysaccharide dextran are critical for preserving dried protein samples by forming rigid a glass that protects entrapped protein molecules. Stably dried proteins are important for maintaining critical information in clinical samples like blood serum. However, we found that dextran reacts with serum proteins during storage, producing high-molecular weight Amadori-product conjugates. These conjugates appeared in a matter of days or weeks when stored at elevated temperatures (37° or 45°C), but also appeared on a timescale of months when stored at room temperature. We synthesized a less reactive dextranol by reducing dextran’s anomeric carbon from an aldehyde to an alcohol. Serum samples dried in a dextranol-based matrix protected the serum proteins from forming high-molecular weight conjugates. The levels of four cancer-related serum biomarkers (prostate specific antigen, neuropiln-1, osteopontin, and metalloproteinase 7) decreased, as measured by immunoassay, when serum samples were stored for one to two weeks in dextran-based matrix. Switching to a dextran-based lyoprotection matrix slightly reduced the damage to osteopontin and completely stopped any detectable damage during storage in the other three biomarkers when for a period of two weeks at 45°C. Dextranol offers a small and easy modification to dextran that significantly improves the molecule’s function as a lyoprotectant by eliminating the potential for damaging protein-polysacharide conjugation.

A Novel and Cost Effective Method of Removing Excess Albumin from Plasma/Serum Samples and Its Impacts on LC-MS/MS Bioanalysis of Therapeutic Proteins

2014 ◽  
Vol 86 (16) ◽  
pp. 8336-8343 ◽  
Author(s):  
Guowen Liu ◽  
Yue Zhao ◽  
Aida Angeles ◽  
Lora L. Hamuro ◽  
Mark E. Arnold ◽  
...  
Keyword(s):  
Therapeutic Proteins ◽  
Cost Effective ◽  
Serum Samples ◽  
Cost Effective Method

scholarly journals A Novel, High-Throughput Workflow for Discovery and Identification of Serum Carrier Protein-Bound Peptide Biomarker Candidates in Ovarian Cancer Samples

2007 ◽  
Vol 53 (6) ◽  
pp. 1067-1074 ◽  
Author(s):  
Mary F Lopez ◽  
Alvydas Mikulskis ◽  
Scott Kuzdzal ◽  
Eva Golenko ◽  
Emanuel F Petricoin ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Serum Proteins ◽  
Early Stage ◽  
Direct Sequencing ◽  
Stage I ◽  
Clinical Samples ◽  
Carrier Protein ◽  
Serum Samples ◽  
Workflow System ◽  
Affinity Enrichment

Abstract Background: Most cases of ovarian cancer are detected at later stages when the 5-year survival is ∼15%, but 5-year survival approaches 90% when the cancer is detected early (stage I). To use mass spectrometry (MS) of serum proteins for early detection, a seamless workflow is needed that provides an opportunity for rapid profiling along with direct identification of the underpinning ions. Methods: We used carrier protein–bound affinity enrichment of serum samples directly coupled with MALDI orthagonal TOF MS profiling to rapidly search for potential ion signatures that contained discriminatory power. These ions were subsequently directly subjected to tandem MS for sequence identification. Results: We discovered several biomarker panels that enabled differentiation of stage I ovarian cancer from unaffected (age-matched) patients with no evidence of ovarian cancer, with positive results in >93% of samples from patients with disease-negative results and in 97% of disease-free controls. The carrier protein–based approach identified additional protein fragments, many from low-abundance proteins or proteins not previously seen in serum. Conclusions: This workflow system using a highly reproducible, high-resolution MALDI-TOF platform enables rapid enrichment and profiling of large numbers of clinical samples for discovery of ion signatures and integration of direct sequencing and identification of the ions without need for additional offline, time-consuming purification strategies.

scholarly journals Pneumococcal Serotype Identification by Capsular Sequence Typing (CST): A Modified Novel Approach for Serotyping Directly in Clinical Samples

Diagnostics ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 2353
Author(s):  
Nektarios Marmaras ◽  
Athanasia Xirogianni ◽  
Anastasia Papandreou ◽  
Efthymia Petinaki ◽  
Vana Papaevangelou ◽  
...  
Keyword(s):  
Vaccination Strategy ◽  
Cost Effective ◽  
Clinical Samples ◽  
Pneumococcal Serotypes ◽  
Conjugate Vaccines ◽  
Vaccination Programs ◽  
Epidemiological Monitoring ◽  
Novel Approach ◽  
Cost Effective Method ◽  
Serotype Identification

As almost 60–70% of Invasive Pneumococcal Disease (IPD) is identified by nonculture methods in Greece, serotyping is of high importance for the better monitoring of pneumococcal serotypes due to the availability of conjugate vaccines. The aim of the study was the modification and direct application of the Capsular Sequence Typing (CST) assay in clinical samples in order to serotype Streptococcus pneumoniae culture-negative, Polymerase Chain Reaction (PCR_-positive samples, followed by CST group specific single-tube PCR assays. A two-step PCR modified assay was applied on a total of 306 samples (such as CSF, blood, pleural and middle ear fluids, isolates) obtained from 283 patients with IPD. The overall performance permits a rapid, accurate and cost-effective method for nonculture pneumococcal serotyping. As the management of IPD is closely related to the continuous monitoring of pneumococcal serotypes, the proposed approach proved to be a valuable tool for the typing and epidemiological monitoring of S. pneumoniae, for the evaluation of the overall impact of vaccination programs in the era of pneumococcal conjugate vaccines, in order to initiate the appropriate vaccination strategy.

scholarly journals Electrophoretic Profile of Serum Proteins Using Capillary Technique in Patients Attending the Douala General Hospital, Cameroon

2018 ◽  
Vol 6 (2) ◽  
pp. 50-55
Author(s):  
Judith Gwladys Ekwe Priso ◽  
Jean Pierre Nda Mefo’o ◽  
Cécile Okalla Ebongue ◽  
Eveline Ngouadjeu Dongho Tsakeu ◽  
Catherine Akono Ndi ◽  
...  
Keyword(s):  
Capillary Electrophoresis ◽  
General Hospital ◽  
Gel Electrophoresis ◽  
Serum Proteins ◽  
Agarose Gel Electrophoresis ◽  
Serum Samples ◽  
Electrophoretic Profile ◽  
Biuret Method ◽  
Accuracy Of Results ◽  
Separation Of Proteins

Background: Electrophoresis of serum proteins is an orientation examination routinely used in clinical practice. For a few years, agarose gel electrophoresis has tended to be replaced with capillary electrophoresis owing to an increase in the accuracy of results. However, this technique is uncommon and is not widely used in Cameroon. Objectives: The research aimed at studying the electrophoretic profile of serum proteins using capillary technique among patients attending the Douala General Hospital, Cameroon. Methods: Capillary electrophoresis was used to carry out tests on blood samples from any inpatients and outpatients and fasting for 8-12 hours. Capillary electrophoresis of serum samples was used for the separation of proteins into six fractions and the total protidemia of each serum samples was determined using the Biuret method. Results were interpreted by observing the shape of curves and quantitative variations in each fraction of the different serum proteins. Results: A total of 311 patients participated in the study. The sampled population aged 50±18 years on average and consisted of 55.3% men and 44.7% women. All capillary electrophoresis profiles presented six protein fractions, namely, albumin, alpha (1 and 2), beta (1 and 2) and gamma globulins. Pathological disorders were diagnosed in 290 patients and 21 patients had normal results. Inflammatory syndromes accounted for 63.34% and monoclonal gammopathies for 10.29% the main pathological disorder identified. Conclusion: Capillary electrophoresis provides a more precise identification of biological syndromes and clear distinction of the six fractions of each protein. Monoclonal profiles and inflammatory syndromes were well detected. A prevalence of 10.29% was determined for gammopathies.

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