Datura stramonium Lectin (DSL) Production Service at CD BioGlyco

Lectins are glycan-binding proteins that play a very important role in biomolecule recognition, cell communication, immune regulation, and protein transport. Lectins have been characterized in legumes, Solanaceae, etc. CD BioGlyco provides clients with professional Lectin Production Services of Natural Origin such as plant origin, animal origin, and fungal origin. D. stramonium, also known as thorn apple and jimson weed, is an annual tropical herb of the genus Datura in the Solanaceae family. DSL is extracted and purified from this plant of the Solanaceae family, specifically binds chitin, and has antimicrobial and anti-tumor activity. The carbohydrate-binding site of this lectin preferentially recognizes complex N-glycans containing three or more branches of N-acetyllactosamine moieties rather than single N-acetylglucosamine residues. Among three-branch complex N-glycans, DSL preferentially binds to C-2,6-branched rather than C-2,4-branched N-glycans. Studies have found that β1-4-linked N-acetyllactosamine or N-acetylglucosamine oligomers inhibit hemagglutination activity.

Fig.1 DSL production service. (CD BioGlyco)

• Extraction and Purification of DSL

D. stramonium seed is ground in a seed mill and extracted with phosphate-buffered saline. The obtained crude cell-free extract is centrifuged, the supernatant is taken, and the precipitate is collected after treatment with acetone. The precipitate is resuspended in phosphate buffer, dialyzed, and purified by affinity chromatography.

• Determination of Molecular Mass

Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is performed using a precast homogeneous gel, and proteins in the gel are stained with Coomassie brilliant blue (CBB). The molecular weight of DSL is estimated at the constructed calibration curve of the marker.

• Analysis of N-Terminal Amino Acid Sequence

To determine the N-terminal sequence of DSL subunits, each subunit band electroblotted onto a polyvinylidene difluoride (PVDF) membrane is excised and analyzed with a protein sequencer.

• Agglutination Assay

Agglutination is one of the easiest immunological tests to perform and is used to detect antigens or antibodies. We test for hemagglutination by serially diluting the sample to be tested in phosphate buffer and then mixing it with an equal volume of rabbit red blood cell suspension.

• Determination of Lectin Activities
  • Blood group specificity
  • Effect of pH on DSL activity
  • Effect of temperature on DSL activity
  • Determination of the antifungal activity of DSL
  • Determination of the antiproliferative activity of DSL

We also provide clients with a variety of DSL products to choose from, including but not limited to:

Publication

Highlights

• We use affinity chromatography to purify the lectins and separate the different fractions.
• We provide global clients with extraction, purification, characterization, bioactivity testing, sequencing, and other services of DSL to support your scientific research.
• DSL is not only used in lectin microarrays for glycomic analysis but is also used in glycoproteomics studies in combination with mass spectrometry techniques.

CD BioGlyco provides a broad range of Lectin Production Services to identify specific carbohydrates to support your glycomics and glycoproteomics research. Please feel free to contact us if you have any lectin production needs.