Materials and Reagents

1. Materials
   1.96 well microtiter plates (V bottom) (Corning, catalog number: 3897 )
   2.Tips for multichannel pipette (Gilson, model: D10 , D200 and D1000 )
   3.Centrifuge tubes (SARSTEDT AG & Co, catalog number: 72.690 )
   
   
2. Reagents
   
   1. Viral antigen
      
      1. H1N1
         A/California/07/2009
      2. H3N2
         A/Brisbane/10/2007
         Note: Viruses were amplified in embryonated eggs. Refer to Manual for the Laboratory Diagnosis and Virological Surveillance of Influenza, WHO.
         
   2. Erythrocytes
      Turkey red blood cells (RBCs)
      Note: Both in-house RBCs and commercial RBCs are usable.
      
   3. Antibody
      Human monoclonal antibodies CT146, CT147, CT149, CT164 and CT166
      Notes:
      
      1. Maximum concentration of antibody is 20 μg/ml.
         
      2. Expressed in CHO cells.
         
      3. Purified according to manufacturer’s instruction [HiTrap^TM MabSelect SuRe (GE Healthcare, catalog number: 11-0034-93 )].
   4. Other reagents
      1. Receptor destroying enzyme (RDE) (DENKA SEIKEN CO., catalog number: 370013 )
         
      2. Phosphate buffered saline (1x PBS) (pH 7.2) (Sigma-Aldrich, catalog number: P4417 )
         
      3. Physiological saline (0.85% NaCl) (Sigma-Aldrich, catalog number: S9888 )

Equipment

1. Hemocytometer (Hausser Scientific, catalog number: 1492 )
   
2. Multichannel pipette (Gilson, model: PIPETMAN Neo^® Multichannel )
   
3. Centrifuge (Beckman Coulter, model: Allegra X-15R )
   
4. Water bath (JULABO GmbH, model: MB13 )
   
5. Inverted routine microscopy (Nikon Instruments Inc., model: TS-100 )
   

Procedure