Efficient purification and preconcentration of erythropoietin in human urine by reusable immunoaffinity column

Mi, Jiebo; Wang, Shan; Ding, Xiaojie; Guo, Zhiqiang; Zhao, Meiping; Chang, Wen‐Bao

doi:10.1016/j.jchromb.2006.05.037

Cited by 12 publications

(2 citation statements)

References 25 publications

(20 reference statements)

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“…Lonnberg et al (2010) has shown that the anti-EPO monolith column is able to remove 99.7% of undesirable proteins in urine samples. The advantage of anti-EPO monolith column includes its high specificity and selectivity antibody (Mi et al, 2006). Although all urine samples were pre-spiked with the same amount of EPO, the anti-EPO monolith column was able to recover more uhEPO from urine than the anti-EPO micro well plate, based on the intensity of the bands formed.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Methods for Purifying Erythropoietin from Urine using Immunoaffinity Applications

Kwan¹,

Baharudin²,

Salim³

et al. 2015

International J. of Biological Chemistry

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Erythropoietin (EPO), a hormone that regulates the synthesis of red blood cells, is frequently abused by athletes. Sodium dodecyl sulphate poly acrylamide gel electrophoresis (SDS-PAGE) is an essential analytical technique in all anti-doping laboratories in order to detect the abuse of EPO. An immunoaffinity purification step is now considered essential for the pre-treatment of urine samples to isolate EPO prior to gel electrophoresis. In this study, we have compared the performance of two immunoaffinity purification techniques in EPO anti-doping analysis, i.e., the anti-EPO micro well plate and anti-EPO monolith column. The anti-EPO monolith column is efficient in removing undesirable proteins except for the Bovine Serum Albumin (BSA) as seen on SDS-PAGE. The BSA was eventually removed from the protocol and the undesirable protein band was eliminated without affecting the performance of the method. Throughout the study, the anti-EPO monolith column emerged as a better option, as it provided a higher sensitivity and higher throughput analysis when compared to the anti-EPO micro well plate. The anti-EPO monolith column has shown consistent results with the EPO recovery rate of 72%, while the limit of detection is as low as 0.5 mIU mLG 1 .

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Section: Discussionmentioning

confidence: 99%

Evaluation of Methods for Purifying Erythropoietin from Urine using Immunoaffinity Applications

Kwan¹,

Baharudin²,

Salim³

et al. 2015

International J. of Biological Chemistry

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“…In many cases the techniques described utilize paramagnetic beads with an anti-EPO antibody attached to the surface. Another option is for the anti-EPO antibody to be held on a monolithic polymer support inside a disposable holder [20] [21] or on a chromatographic gel bed [22].…”

Section: Introductionmentioning

confidence: 99%

Improved Recovery of Erythropoietin and Darbepoetin from Equine Plasma by the Application of a Wheat Germ Agglutinin Mediated Pre-Extraction Prior to Immunoaffinity Chromatography

Stanley¹,

Chua²

2014

ABB

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We describe a two-step method that uses wheat germ agglutinin immobilized on Sepharose gel followed by immunoaffinity chromatography (IAC) to extract recombinant human erythropoietin and Darbepoetin from equine plasma. Lectin affinity chromatography was shown to be an effective approach for isolating these epoetins from plasma and in combination with IAC; this method gave superior recovery when compared to the use of the latter technique alone. Moreover, due to the ease with which it can be scaled up, it is particularly well suited for pre-concentrating larger volumes of samples prior to IAC and this provides a facile way of improving the overall sensitivity with which these foreign proteins can be detected in equine plasma.

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Survey of the year 2006 commercial optical biosensor literature

Rich

Myszka

2007

J of Molecular Recognition

109

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We identified 1219 articles published in 2006 that described work performed using commercial optical biosensor platforms. It is interesting to witness how the biosensor market is maturing with an increased number of instrument manufacturers offering a wider variety of platforms. However, it is clear from a review of the results presented that the advances in technology are outpacing the skill level of the average biosensor user. While we can track a gradual improvement in the quality of the published work, we clearly have a long way to go before we capitalize on the full potential of biosensor technology. To illustrate what is right with the biosensor literature, we highlight the work of 10 groups who have their eye on the ball. To help out the rest of us who have the lights on but nobody home, we use the literature to address common myths about biosensor technology.

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Efficient purification and preconcentration of erythropoietin in human urine by reusable immunoaffinity column

Cited by 12 publications

References 25 publications

Evaluation of Methods for Purifying Erythropoietin from Urine using Immunoaffinity Applications

Evaluation of Methods for Purifying Erythropoietin from Urine using Immunoaffinity Applications

Improved Recovery of Erythropoietin and Darbepoetin from Equine Plasma by the Application of a Wheat Germ Agglutinin Mediated Pre-Extraction Prior to Immunoaffinity Chromatography

Survey of the year 2006 commercial optical biosensor literature

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