Difference of cellular affinity against SBA-lectin

• Materials and methods
• G-Coat dishes modified with α-1,6-galactose
• SBA (Soybean agglutinin) is a plantal glycoprotein composed of four subunits having
• BIBa specific binding domain to N-acetylgalactosamine and galactose.
• Venous blood of healthy adults was used to collect blood cells except hematopoietic BIBstem cells collecting from cord blood.
• Identification of blood cell types was carried out by flow cytometry using monoclonal BIBantibodies (Beckton Dickinson).

• Results
• The order of cell adhesion based on the affinity with lectins:
• BIBErythroid, granulocytes ＞ lymphocytes ＞NK celis
• In the case of applying less than 100 μg/mL of SBA, erythrocytes preferentially BIBadhered to the dishes. Thereby leukocytes were isolated from erythrocytes as
• BIBan unattached cell fraction.
• The adhesion of the immature stem cells was 21% even if the dishes were treated BIBby over 800 μg/mL of SBA.

Enrichment of NK cells using ECL-lectin (Negative selection)

• Materials and methods
• The dishes (Φ: 35mm) were modified with G-coat involving galactose and glucose, BIBand then treated with galacto-specific ECL (10 ng/mL) at room for 60 min.
• Peripheral blood mononuclear cells (PBMCs) were enriched by 1.077 g/mL single-BIBdensity gradient centrifugation.
• PBMCs (2x10) including NK cells were resuspended in dPBS(-) with 0.2 wt% of BIBBSA.
• The suspended cells were seeded on the ECL-treated dishes, and then incubated at BIB37℃ for 30 min.
• Nonadherent cells in supernatant were recovered from the dishes, and then NK cells BIBand lymphocytes in the recovery were identified by Flow cytometry using
• BIBmonoclonal antibodies.

• Results
• The loss of NK cells was suppressed by diminishing galactose ratio on the dishes, BIBto lead to more selective collection of NK cells.
• The adjustment of buffer or incubation conditions may improve the efficiency of NK BIBcell enrichment.

Basic functions of lectin
Efficacy of lectin
Practicability
Ripple effect