A recombinant Fab fragment-based electrochemical immunosensor for the determination of testosterone in bovine urine

Lu, Huihui; Kreuzer, Mark P.; Takkinen, Kristiina; Guilbault, George G.

doi:10.1016/j.bios.2006.08.002

Cited by 43 publications

(24 citation statements)

References 34 publications

(30 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Testosterone was easily identified and quantified. Despite the small crossreactivity reported for epitestosterone and the Fab fragment [36], epitestosterone was not detected in PF-MEKC analysis.…”

Section: Determination Of Testosterone In Male Urine Samplesmentioning

confidence: 70%

“…In the off-line construction, androstenedione and methyltestosterone could be used as external standards in PF-MEKC analyses after IA-SPE. The resolving power of PF-MEKC was also beneficial in view of the minor cross-reactivity between the antitestosterone Fab fragment and epitestosterone [36].…”

Section: Discussionmentioning

confidence: 99%

“…The SDS alone does not provide resolution between structurally similar steroids [16], and we could not use an absorbing resolving component, such as taurocholic acid [23], in the micellar phase since they obscured testosterone signal at 247 nm. In the analysis of urine samples by the hyphenated IA-SPE-PF-MEKC system, the cross-reactivity reported for the Fab fragment and epitestosterone [36] could not have been compensated with MEKC. Hence, we focused our further studies on off-line IA-SPE and PF-MEKC.…”

Section: Hyphenated Ia-spe-pf-mekc Analysis Of Testosteronementioning

confidence: 99%

“…A microscale column filled with the immunosorbent was employed in the extractions, and the eluted fractions were analyzed by a selective PF-MEKC method. In contrast to the hyphenated IA-SPE-PF-MEKC method, the resolving power of the off-line PF-MEKC method could be optimized to enable the use of external standards and to compensate the minor crossreactivity (7.2%) of the antitestosterone Fab fragment and epitestosterone [36].…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Microscale immunoaffinity SPE and MEKC in fast determination of testosterone in male urine

et al. 2007

Self Cite

View full text Add to dashboard Cite

Conventional methods for the determination of testosterone in body fluids typically suffer from poor recovery, lack of specificity, complex sample pretreatment, or the need for derivatization. Here, a simple, specific, and fast analysis method for testosterone was developed, with a methodology based on testosterone-specific immunoaffinity SPE (IA-SPE) and subsequent analysis by partial filling MEKC (PF-MEKC). An immunosorbent consisting of a recombinant antitestosterone Fab fragment covalently attached to activated Sepharose was prepared. IA-SPE and PF-MEKC were set up in hyphenated and off-line constructions, and the applicability of the two constructions in analysis of testosterone in male urine was investigated. The results obtained with the hyphenated construction proved to be only indicative of the presence of testosterone. The off-line IA-SPE and PF-MEKC construction, however, was successfully used in the determination of free testosterone in male urine samples after enzymatic hydrolysis of the glucuronide conjugates. Except for the hydrolysis reaction, no sample pretreatment was required. After hydrolysis, the overall analysis time per sample was only 14 min. The off-line IA-SPE and PF-MEKC method proved to be robust, sensitive (LOQ 35 mg/L), and specific, enabling separation of testosterone from four related steroids. Thus, it provides attractive features when compared to traditional methods for determination of testosterone in male urine.

show abstract

Section: Determination Of Testosterone In Male Urine Samplesmentioning

confidence: 70%

Section: Discussionmentioning

confidence: 99%

Section: Hyphenated Ia-spe-pf-mekc Analysis Of Testosteronementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Microscale immunoaffinity SPE and MEKC in fast determination of testosterone in male urine

et al. 2007

Self Cite

View full text Add to dashboard Cite

show abstract

“…These analytical methods provide quantitative and confirmatory results with high sensitivity and selectivity, but require some expensive instrumentation and suffer from considerable time delays between sampling and obtaining the results. These disadvantages limit their daily routine use in laboratory environments [6]. Electrochemical techniques using modified electrodes have proved to be a sensitive and versatile method for the determination of analytes undergoing biologically important reactions of oxidation and/or reduction, including isolated drugs and related molecules in pharmaceutical dosage forms.…”

Section: Introductionmentioning

confidence: 99%

Electrochemical detection in vitro and electron transfer mechanism of testosterone using a modified electrode with a cobalt oxide film

Moura

Moraes

Santos

et al. 2014

Sensors and Actuators B: Chemical

View full text Add to dashboard Cite

A biosensor platform based on amine functionalized conjugated benzenediamine‐benzodithiophene polymer for testosterone analysis

Bulut

Sanli

Cevher

et al. 2020

J of Applied Polymer Sci

View full text Add to dashboard Cite

A novel benzenediamine-benzodithiophene polymer is synthesized for use in biosensor fabrication for the detection of testosterone. The sensory platform is constructed via drop coating on a screen-printed carbon electrode, using poly(benzenediamine-Bis[(2-ethylhexyl)oxy]benzodithiophene) (pBDBT) as the polymer layer. Testosterone antibodies are immobilized on the polymercoated electrode surface via glutaraldehyde, which binds to the surface through the amino functional groups on the polymer backbone. The changes in the surface features due to testosterone binding are investigated via electrochemical techniques such as differential pulse voltammetry, cyclic voltammetry, and electrochemical impedance spectrometry as well as contact angle measurements. Surface morphology of the modified electrodes is characterized by atomic force microscopy. The linear range and limit of detection of the sensor are calculated. Impact of possible interfering compounds is investigated. Furthermore, the sensory platform is utilized for testosterone analysis in synthetic biological fluids.

show abstract

A recombinant Fab fragment-based electrochemical immunosensor for the determination of testosterone in bovine urine

Cited by 43 publications

References 34 publications

Microscale immunoaffinity SPE and MEKC in fast determination of testosterone in male urine

Microscale immunoaffinity SPE and MEKC in fast determination of testosterone in male urine

Electrochemical detection in vitro and electron transfer mechanism of testosterone using a modified electrode with a cobalt oxide film

A biosensor platform based on amine functionalized conjugated benzenediamine‐benzodithiophene polymer for testosterone analysis

Contact Info

Product

Resources

About