The Molecular Technologies facility provides access to expertise and modern technologies for the generation and expansion of small compound libraries, the development of analogue series, and the identification, characterisation and quantification of specific compounds and natural extracts through its broad range of analytical expertise.

Equipment:

• State-of-the-art technologies for chemical synthesis (Chemspeed robotic and microwave synthesis platforms)
• Semi-preparatory and preparatory chromatography (including TLC, HPLC, CCC)
• Associated detection methods (UV/VIS, fluorescence, MS)
• NMR technologies and in-silico platforms for the identification of molecules and the elucidation of their 3D structure.

The facilities capabilities include:

Chemical library management:

• Compound management and hosting
• Miniaturization
• Natural and synthetic collections
• Library generation based on specific chemotypes

For small and natural compounds:

• Identification and isolation from complex matrices
• Natural and simplified ADME model
• Chemical modifications
• Bioactivation/inactivation models
• In-silico modelling and design

Analysis of small and large molecules:

• Biomarker ID and quantification in products (QC)
• Biomarker ID and quantification in complex matrices (PK, Clinical studies)
• Characterisation and quantification of specific analytes
• Purity determination
• Method development and validation

Contact:
Library and compound synthesis
Dr Edwin MMutlane
Molecular Technologies
CSIR Biosciences
Tel: 012 841 4282
Email:

Compound analysis
Dr Paul Steenkamp
Molecular Technologies
CSIR Biosciences
Tel: 012 841 3260
Email:

The pharmacology and bioassay facility designs, develops and validates pharmacological assays for drug benefit and risk assessment, which are applied in biomolecular screening campaigns of small to medium sized chemical and extract libraries.

Equipment:

• State-of-the-art liquid handling and high-throughput robotic assay facilities
• High-end spectrophometry for absorbance/fluorescence measurements
• Realtime PCR
• Agilent Bioanalyzer analysis for RNA, DNA and protein integrity

Services:

In vitro models for drug benefit and risk assessment include:

Pathogens and infectious disease

• Antiviral screens including HIV pseudovirus
• Malaria (conventional and transmission blocking screens)
• Antimicrobial screens
• Antifungal screens
• Drug signature profiling

 

Mammalian cell tissue culture

• Cancer and oncology screens
• Drug permeability
• Drug cytotoxicity screening
• Drug signature profiling

The facility also develops and applies assays for cosmeceuticals testing. In vitro models include:

Enzyme and cell-based assays

• Elastase, collagenase, hyaluronidase screening for activity inhibition
• Elastin and collagen screening for protein production

Targeted and whole genome assays

• Cell-based TaqMan and microarray expression profiling of molecular biomarkers specific to youthful skin and ageing

Contact:
Ms Natasha Kolesnikova / Dr Malefa Tselanyane
Biomedical Technologies
Tel: 012 841 4735 / 012 841 4045
Email: /

The biomarker discovery/validation and biomolecule analysis facility characterises and quantifies specific compounds through a broad range of analytical services.

Equipment:

Electrophoresis
Ettan IPGphor I/II IEF (GE Amersham)
Rotofor Cell (Bio-Rad)
Dalt II Separation Unit (GE Amersham)
Protean Plus DodecaCell (Bio-Rad)
Ettan SE600 Ruby gel electrophoresis system with auto gel stainer
Ettan DALTtwelve, running 12 large format gels in parallel

Imaging
PharosFX (Bio-Rad)
EXQuest automated spotcutter (Bio-Rad)
PDQuest (Bio-Rad)
Image Master 2D scanner and software

HPLC
1100 nanoHPLC (Agilent Technologies)

Mass spectrometry
MAPII AutoPrep MALDI Robot system (Bruker Daltonics)
Bruker autoflex® Daltonics MALDI-TOF mass spectrometer with post-source decay
QStar Elite (ESI/MALDI) (Applied Biosystems)
Qtrap (ESI) (Applied Biosystems)

Protein-interaction
Biocore 3000 (GE Amersham)

Services:

Proteomics services are available to internal and external customers. The following services are available:

Biomarker & Drug-target discovery and validation

Gel-based proteomics

• • Fractionation: 1D / 2D PAGE;
  • Protein visualisation / isolation: gel staining (Coommassie, Silver, SYPRO-Ruby, Cy dye, and Flamingo)
  • Gel imaging (fluorescence, colorimetric and radioisotopic) (Biorad PharosFX)
  • High-throughput spot picking (Biorad ExQuest); and
  • 1D / 2D PAGE band/spot identification/confirmation  (Dionex Ultimate 3000 and AB Sciex QStar Elite).

Solution-based (gel-free) proteomics

• Off-line and on-line fractionation:

• In-solution peptide/protein IEF (Biorad MicroRotofor)
• 2D HPLC (Dionex Ultimate 3000 nanoRSLC

• Shotgun and targeted biomarker/drug target discovery  (AB Sciex QStar Elite and AB Sciex 4000 QTRAP)

• MRM-based During ‘validation’, putative targets are confirmed by a targeted approach where fewer analytes are monitored in a larger sample size. The main aim of this stage is to assess marker sensitivity (likelihood that an affected/diseased sample will test positive). In ‘verification’ the analysis is further extended with 100s of samples analysed in order to examine variations due to environmental, genetic and/or biological factors. Thus, candidate biomarker sensitivity is affirmed while specificity (likelihood that an unaffected/healthy sample will test negative) is gauged.”

• Mass spectrometry (iTRAQTM, peptide identification, peptide sequence identification, glycan analysis, posttranslational modification analysis)
• Protein characterisation (protein-protein / ligand interaction)

Microarrays (cDNA oligo)

•     Sample preparation and hybridisation
•     Conventional and high-resolution scanning*
•     Expression analysis
•     Quantitative Real-Time PCR

Contact information
Dr Stoyan Stoychev

Molecular Technologies

Tel: +27 12 841 2270/3001

Fax: +27 12 841 2388

Email: