Activity of different proteases in a complex mixture and in vitro study of their reciprocal interferences by micellar electrokinetic chromatography

Viglio, Simona; Lupi, A.; Luisetti, Maurizio; Zanaboni, Giuseppe; Cetta, Giuseppe; Iadarola, Paolo

doi:10.1002/(sici)1520-667x(2000)12:5<302::aid-mcs4>3.0.co;2-#

Cited by 4 publications

(2 citation statements)

References 16 publications

(1 reference statement)

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“…Recent advances in CE have indicated that this technique has become the method of choice for determining numerous proteolytic activities in vitro and in vivo [18,19,21,[26][27][28]. To investigate whether the applicability of CE in free solution or MEKC can be further extended to measure TPP-I activity, human skin fibroblasts cultures were considered as a source of this proteinase.…”

Section: Determination Of Tpp-i Activity On Fibroblasts Of Healthy Sumentioning

confidence: 99%

“…Although presently used, both the above-cited methods are still rather laborious and time-consuming. In this respect, because capillary electrophoresis (CE) has proven to be an excellent tool for monitoring activities of serine-and metallo-proteinases [18][19][20][21], we explored the possibility of using this technique to assay for TPP-I. This paper describes (i) a systematic investigation carried out with capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC) to optimize the experimental conditions for monitoring TPP-I activity using a synthetic peptide, (ii) the determination of TPP-I activity using various human and animal samples as sources of this ezyme, and (iii) the determination of TPP-I activity in human specimens obtained from patients affected by LINCL.…”

Section: Introductionmentioning

confidence: 99%

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Diagnosis of late-infantile neuronal ceroid lipofuscinosis: A new sensitive method to assay lysosomal pepstatin-insensitive proteinase activity in human and animal specimens by capillary electrophoresis

Viglio¹,

Marchi²,

Wisniewski³

et al. 2001

Electrophoresis

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Batten disease, or human late-infantile neuronal ceroid lipofuscinosis (LINCL) is a familiar progressive degenerative disease affecting children, caused by a deficiency of a lysosomal proteinase (tripeptidyl peptidase I, TPP-I) and characterized by the accumulation of autofluorescent storage bodies in the brain and other tissues of the body. Current methodology used to diagnose this disease needs to be improved in order to have less invasive techniques with higher resolution and shorter assay time. In this report, we discuss the potential merits of micellar electrokinetic chromatography as an excellent tool that requires minute samples but offers high resolution and a short running time for monitoring TPP-I activity in human and animal specimens.

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Section: Determination Of Tpp-i Activity On Fibroblasts Of Healthy Sumentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Diagnosis of late-infantile neuronal ceroid lipofuscinosis: A new sensitive method to assay lysosomal pepstatin-insensitive proteinase activity in human and animal specimens by capillary electrophoresis

Viglio¹,

Marchi²,

Wisniewski³

et al. 2001

Electrophoresis

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Microscale separation methods for enzyme kinetics assays

Křížek

Kubíčková

2012

Anal Bioanal Chem

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Miniaturization continues to be one of the leading trends in analytical chemistry and one that brings advantages that can be particularly beneficial in biochemical research. Use of a miniaturized scale enables efficient analysis in a short time and requires very small amounts of samples, solvents, and reagents. This can result in a remarkable decrease in costs of enzyme kinetics studies, especially when expensive or rare enzymes and/or substrates are involved. Free zone electrophoresis is without a doubt the most common microscale separation technique for capillary and on-chip enzyme assays. Progress and applications in this field are reviewed frequently whereas other modes of separation, although successfully applied, receive only marginal interest in such publications. This review summarizes applications of less common modes of separation in capillary or chip formats, namely micellar electrokinetic chromatography, liquid chromatography, gel electrophoresis, isoelectric focusing, and isotachophoresis. Because these techniques are based on separation mechanisms different from those of free zone electrophoresis, they can be, and have been, successfully used in cases where zone electrophoresis fails. Advantages and drawbacks of these alternative separation techniques are discussed, as also are the difficulties encountered most often and solutions proposed by different research groups.

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Determination of enzymatic activity by capillary electrophoresis

Glatz

2006

Journal of Chromatography B

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Activity of different proteases in a complex mixture and in vitro study of their reciprocal interferences by micellar electrokinetic chromatography

Cited by 4 publications

References 16 publications

Diagnosis of late-infantile neuronal ceroid lipofuscinosis: A new sensitive method to assay lysosomal pepstatin-insensitive proteinase activity in human and animal specimens by capillary electrophoresis

Diagnosis of late-infantile neuronal ceroid lipofuscinosis: A new sensitive method to assay lysosomal pepstatin-insensitive proteinase activity in human and animal specimens by capillary electrophoresis

Microscale separation methods for enzyme kinetics assays

Determination of enzymatic activity by capillary electrophoresis

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