Applications notes

CARBOHYDRATES

69

Application

ANTIBODY GLYCOSYLATION

 

Separation

CE-LIF

λ

488

Description

Glycan characterization of therapeutic proteins is of utmost importance due to the role of carbohydrates in protein stability, half-life, efficacy and mechanism of action. This application note presents the on-line CE-LIF-MS technology for the direct identification of N-linked glycan profiling of rMAbs. CE-MS technology with on-line LIF detection has been developed to provide accurate mass information on major and minor peaks observed in the routine CE-LIF assay.

70

Application

ANTIBODY GLYCOSYLATION

Separation

µLC LIF MS

λ

325

Description

This application note describes the characterisation of carbohydrates structures on therapeutic proteins. 2-aminobenzoic acid dye is used for the analysis of mono or oligosaccharides with a 325nm HeCd laser in HPLC.

2.003

Application

POLYSACCHARIDES

Separation

CE

λ

480

Description

9-Aminopyrene-1,4,6-trisulfonate (APTS) is dye that is frequently used for the analysis of mono or oligosaccharides. The labeling of sugars involves a reductive amination of the reductive function of the mono or oligosaccharides followed by reaction with the dye. APTS is routinely used in Capillary Electrophoresis separation. In this note, we analyze oligosaccharides labeled with APTS with a 480nm LED.

2.004

Application

POLYSACCHARIDES

Separation

µLC

λ

365

Description

Therapeutic Antibodies is a category of therapeutics that plays an important role in controlling many diseases such as cancer, allergy, inflammation, infectious or autoimmune diseases. The characterization of carbohydrates structures on therapeutic proteins is of utmost importance due to their role in clearance and mechanism action. The main techniques for carbohydrates analysis involve high-performance liquid chromatography (HPLC), mass spectrometry (MS) and capillary electrophoresis (CE). 2-aminobenzoic acid (2AB) is a dye frequently used for the analysis of mono or oligosaccharides with HPLC.

NUCLEIC ACIDS

71

Application

DNA SIZE FRACTIONATION WITH µLAS TECHNOLOGY

 

Separation

CE-LIF

λ

488

Description

This application note describes how the µLAS technology is used to fractionate DNA with the Agilent 7100 Capillary Electrophoresis system coupled to Picometrics Laser Induced Fluorescence Detector.

2.005

Application

APTAMERS

Separation

CE

λ

210/480

Description

This application note describes an alternative method of the selection of aptamers using CE-UV-LEDIF detection/non-SELEX.

2.006

Application

APTAMER COLLECTION METHOD

 

Separation

CE

λ

210/480

Description

In this application note, we optimised the collection parameters of aptamers on a standard mixture to validate the collection method in a small range of migration times to get the slower koff.

2.009

Application

DNA SIZING DOWN TO 1 FG/µL SENSITIVITY WITH MULTIPLE INJECTIONS

 

Separation

CE

λ

480

Description

This application note describes a multiple injection method to enhance detection sensitivity of DNA down to 1 fg/µL with µLAS technology.

 

NEUROCHEMISTRY

60 A

Application

AMINO ACIDS AND CATHECHOLAMINES

Separation

FastLC

λ

410

Description

This application note presents the comparison between UPLC/LIF, capillary LC/LIF and CE/LIF analysis of 20 primary amino acids and catecholamines labeled with NDA in microdialysates. It permits to do the best compromise for the neuroscientist needs. The three methods were optimisated to show the main amino acids and catecholamines in neurobiological samples, but they are adjustable, depending on what amino acids the neurochemist need for his study. We used NDA (naphthalene-2,3-dicarboxyaldehyde) because it perfectly allows derivatizing primary amines containing molecules at concentrations as low as 100 picomolar.

60 B

Application

AMINO ACIDS AND CATHECHOLAMINES

Separation

CapLC

λ

410

Description

This application note presents the comparison between UPLC/LIF, capillary LC/LIF and CE/LIF analysis of 20 primary amino acids and catecholamines labeled with NDA in microdialysates. It permits to do the best compromise for the neuroscientist needs. The three methods were optimisated to show the main amino acids and catecholamines in neurobiological samples, but they are adjustable, depending on what amino acids the neurochemist need for his study. We used NDA (naphthalene-2,3-dicarboxyaldehyde) because it perfectly allows derivatizing primary amines containing molecules at concentrations as low as 100 picomolar.

60 C

Application

AMINO ACIDS AND CATHECHOLAMINES

Separation

CE

λ

410

Description

This application note describes a method to simultaneously detect and quantify 21 Amino acids and catecholamines labeled with Naphthalene Dicarboxaldehyde (NDA) in a single injection. The method, developed by Picometrics, couples capillary electrophoresis separation with LIF detection to provide the analyst with a selective and sensitive tool for the analysis of biological samples.

2.002

Application

AMINO ACIDS – CATHECHOLAMINES

Separation

CE/CapLC

λ

450

Description

This application note presents the comparison between CE/L(ed)IF and capillary LC/L(ed)IF analysis of 20 primary amino acids and catecholamines labelled with NDA (naphthalene-2,3-dicarboxyaldehyde).

2.007

Application

AMINO ACIDS: TRYPTOPHAN/TYROSINE/HIAA/HMMA

Separation

CE

λ

275

Description

This application note describes the native fluorescence detection of Tryptophan, Tyrosine, HIAA and HMMA with a ZETALIF LED 275 Detector.

FOOD

65

Application

MYCOTOXINS

Separation

FastLC/CapLC

λ
325
Description

Mycotoxins are toxins that are naturally produced by organisms of the fungus family such as mushrooms, molds and yeasts. These toxins are commonly found in foodstuffs derived from plants, such as cereals, coffee, nuts, fruits, spices and beverages. Many toxins show various toxicological manifestations ; some are teratogenic, mutagenic and/or carcinogenic, and are associated with various diseases. In this application note, we compare the run time and sensitivity for Aflatoxins M2, B2, G2 and Ochratoxin A using a Fast-LC/LIF method and a capillary-LC/LIF method.

DRUGS

66

Application

PROTEIN IgG

Separation

CE-LIF

λ

266/488

Description

This application note describes the separation of Anthracyclines (Doxorubicinol, Doxorubicin and Daunorubicin) using HPLC and Laser Induced Fluorescence Detection.

68

Application

AMPHETAMINES

Separation

CE

λ

410

Description

This note describes the analysis of ten amphetamine derivatives performed by CE-LIF at 410 nm and at very low limits of detection.

2.001

Application

MONONCLONAL ANTIBODIES (mAbs) ANALYSIS

Separation

CE

λ

480/530

Description

Monoclonal antibodies have become a fast growing class of biopharmaceutical products.
To support analytical characterization in process development and quality control of therapeutic antibodies, capillary electrophoresis-sodium dodecyl sulfate (CE-SDS) has been recognized as an important tool in place of SDS-Page because of the ease of use and the ability to automate. In this application note, CE derivatization technique for LED Induced Fluorescence detection have been tested to increase detection sensitivity of proteins with the use of 5-carboxytetramethylrhodamine succinimidyl ester (5-TAMRA.SE) and 3-(2-furoyl)-quinoline-2 carboxaldehyde (FQ) as derivatizing agents.

2.008

Application

MONONCLONAL ANTIBODIES (mAbs) ANALYSIS

Separation

CE

λ

275

Description

Pharmaceutical antibodies are a class of therapeutics that plays an important role in controlling many types of diseases such as cancers, allergy, inflammation, infectious or autoimmune diseases. Monoclonal antibodies have become a major class of biopharmaceutical products. There is an increasing demand in the pharmaceutical industry for tools applicable to large scale production and for product characterization and control of the manufacturing process. To support analytical characterization, process development and quality control of therapeutic antibodies in the biopharmaceutical industry, capillary electrophoresis-sodium dodecyl sulfate (CE-SDS) has been recognized as a significant alternative tool of SDS Page because of its ease of use and ability to automate. This application note describes the analytical methodology for quality control analyses of IgG and their impurities using a 275nm LEDIF detector.

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