A simple semi-automated method of immunoelectrophoresis

Cawley, Leo P.; Schneider, Dezirae; Eberhardt, Lucile; Harrouch, J.; Millsap, G.

doi:10.1016/0009-8981(65)90115-4

Cited by 7 publications

(6 citation statements)

References 4 publications

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“…To improve the speed of the reaction from the slow diffusion step of the RID, Laurell supplied electrophoresis to propel the unknown antibody solution into the gel creating precipitin bands that resembled rockets. This advance allowed the work to be completed in a few of hours [58,59]. Smith and Thompson compared RID results for IgG, IgA and IgM to measurement of the M-protein by densitometric estimations in 186 serum samples from 11 patients with MM or Waldenström macroglobulinemia [60].…”

Section: Radial Immunodiffusionmentioning

confidence: 99%

Challenges of measuring monoclonal proteins in serum

Keren

Schroeder

2016

Clinical Chemistry and Laboratory Medicine (CCLM)

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Abstract:The measurement of monoclonal protein (M-protein) is vital for stratifying risk and following individuals with a variety of monoclonal gammopathies. Direct measurement of the M-protein spike by electrophoresis and immunochemical measurements of specific isotypes or free light chains pairs has provided useful information about the quantity of M-protein. Nonetheless, both traditional electrophoresis and immunochemical methods give poor quantification with M-proteins smaller than 10 g/L (1 g/dL) when in the presence of polyclonal immunoglobulins that co-migrate with the M-protein. In addition, measurements by electrophoresis of M-proteins migrating in the β-and α-regions are contaminated by normal serum proteins in those regions. The most precise electrophoretic method to date for quantification involves exclusion of the polyclonal immunoglobulins by using the tangent skimming method on electropherograms, which provides a 10-fold improvement in precision. So far, however, tangent measurements are limited to γ migrating M-proteins. Another way to improve M-protein measurements is the use of capillary electrophoresis (CE). With CE, one can employ immunosubtraction to select a region of interest in the β region thereby excluding much of the normal proteins from the M-protein measurement. Recent development of an immunochemical method distinguishing heavy/light chain pairs (separately measuring IgGK and IgGL, IgAK and IgAL, and IgMK and IgML) provides measurements that could exclude polyclonal contaminants of the same heavy chain with the uninvolved light chain type. Yet, even heavy/light results contain an immeasurable quantity of polyclonal heavy/ light chains of the involved isotype. Finally, use of liquid chromatography-tandem mass spectrometry (LC-MS/MS) looms on the horizon as a means to provide more consistent and sensitive measurements of M-proteins.

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Section: Radial Immunodiffusionmentioning

confidence: 99%

Challenges of measuring monoclonal proteins in serum

Keren

Schroeder

2016

Clinical Chemistry and Laboratory Medicine (CCLM)

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“…Agar-gel electrophoresis and immunoelectrophoresis were performed at pH 5.5 according to methods previously described [2,3,4,5,6], RAB was prepared by giving each animal 1 ml of equal parts of 2% saline solution of bromelin and Freund's complete adjuvant intra muscularly at 4 sites (hamstrings and shoulders) weekly for 6 weeks. Ten days after the last immunization, sample bleedings were ob tained.…”

Section: Methodsmentioning

confidence: 99%

Immunoelectrophoretic and Serologic Study of the Immunologically Active Components of Bromelin with Rabbit Anti-Bromelin*

Cawley¹,

Schneider²,

Eberhardt³

1966

Vox Sanguinis

Self Cite

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“…After inoculation with antigen and antiserum the sheets are placed on 5" X 7" X 1/8'' lucite plastic trays which are inserted into a specially designed lucite incubation chamber [2,6]. The chamber is made of 1/4" lucite plastic with dimension of 10" X 8" X 8".…”

Section: Methodsmentioning

confidence: 99%

“…Incubation is carried out at room temperature or at 37°C. After precipitin bands are optimally developed (about 24-72 hours) the sheets are placed in film holders (5" X 7"), and soaked first in isotonic saline in photographic tanks (5" X 7") for 24 hours to remove unfixed proteins and buffer salts, followed by distilled water for several hours for removal of saline and then dried in a photographic drying oven [2,6]. The dry sheets 10-B, are stained for protein either with Amido black or thiazine red R. Destaining is performed by rinsing twice in 2% acetic acid.…”

Section: Methodsmentioning

confidence: 99%