Initial Screening of Lectin Property of

Crude Aqueous Plant Seed Extracts

Blood Banking Laboratory
BB 1
Lectins
- a.k.a phytolectin, phytohemagglutinins or phytagglutinins
- are extracts made from seeds of plants which demonstrate hemagglutinating
activity on red cells in vitro
- are proteins that combine specifically with sugars and act as IgM antibodies
- frequently prepared from plants belonging to Family Leguminosae and
Euphorbiaceae
- may also originate from non-plant sources such as animals and microorganisms

Significance of Lectins in blood banking: useful alternative sources of blood typing

sera
Materials/Equipment:
- Gloves - Graduated cylinder
- Centrifuge - Serological pipette
- Wide-mouth bottle (250 mL) - Test tubes
- Beaker - Pasteur pipette
- Nescofilm - Rubber aspirator
- Blender - Test tube rack
- Hot plate - Conical tubes (10 mL)
- Marking pen
Reagents/Samples:
- Normal saline solution (0.85%-0.9% NaCl) in a wash bottle
- 5% red cell suspension (A, B, O, AB)
- Locally available peas, nuts, and beans (e.g. soya bean, lima bean [patani],
mung bean [munggo], peanut)

** Plant voucher specimens must be submitted to the National Museum for proper
identification and authentication.
Procedures:
A. Extract preparation (modified from Das, 2015)
1. Soak 100 g of authenticated plant specimens in 200 mL freshly prepared NSS for 12
hours.
2. Blend the soft seed samples until fine.
3. Transfer blended samples in a beaker and add NSS 3 to 4 times its volume.
4. Mix and then transfer the contents into a 10 mL conical tube.
5. Centrifuge the tube for 10 minutes at 3,400 rpm.
6. Collect the clear supernatant fluid and set it aside
** A 2-fold serial dilution may be done to the supernatant fluid and test the extract against a
batch of 5% red cell suspension.
B. Test for hemagglutinating ability

1. Set up 8 tubes for each blood type and label them accordingly.

Control (CT):
CT1: Anti-A CT3: Anti-AB
CT2: Anti-B CT4: 22% BSA

Test extract (TE):

TE1: Anti-A TE3: Anti-AB
TE2: Anti-B TE4: 22% BSA
2. Proceed as follows:
Control Test extract
Reagent
CT1 CT2 CT3 CT4 TE1 TE2 TE3 TE4
Anti-A 2 drops --- --- --- --- --- --- ---
Anti-B --- 2 drops --- --- --- --- --- ---
Anti-AB --- --- 2 drops --- --- --- --- ---
22% BSA --- --- --- 2 drops --- --- --- ---
Test extract --- --- --- --- 2 drops 2 drops 2 drops 2 drops
A-cell 1 drop --- --- --- 1 drop --- --- ---
B-cell --- 1 drop --- --- --- 1 drop --- ---
AB-cell --- --- 1 drop --- --- --- 1 drop ---
O-cell --- --- --- 1 drop --- --- --- 1 drop
3. Centrifuge the tubes for 15 seconds at 3,400 rpm. Dislodge and interpret.
4. Grade the reaction.
5. Record the results.