CellBind Screen

1
CellBind Screen
Micro-column affinity system
New generation Column technology
2
Methods for antibody detection
Tube test Albumin (BSA) IAT LISS
IAT PEG IAT Column test BioVue ID-sys
tem These tests are based on a visible reaction
caused by agglutination of red cells after adding
anti-human globulin serum
3
Polyspecific Coombs Reagent
Contains Function 1. Anti-IgG detection of
IgG antibodies (37ºC) 2. Anti-C3d detection
of complement dependent IgG or
incomplete IgM antibodies Anti-IgM is not
necessary direct agglutination without AHG
possible in tube- and column agglutination
techniques
4
Advantages of Column technology

• No separate washing steps to be performed
• Easy interpretation of the reactions
• Reactions are stable for a long period
• Possibility of automation

5
New technologies
Solid-phase MTP Solidscreen Capture-R Column
affinity CellBind Screen
6
CellBind columns

• Incubation compartment (LISS medium)
• Density medium
• Affinity matrix

7
Affinity matrix
8
Components of the gel-matrixGel coated with

• Protein G binds IgG antibodies
• Anti-C3d binds complement dependent IgG
  antibodies
• Anti-IgM binds IgM (reactive at 37ºC)

9
Binding of sensitized red blood cells to the gel
matrix ? no agglutination of cells
10
Gel particle red cell visible
using a microscope
Red cell
Gelparticle
11
Microscopic pictures of samples from Cellbind and
Column Agglutination cards
12
Test procedure
40-50 ?L test cells 40-50 ?L patient plasma
or serum (or 20 ?L reagent) Incubate 15
minutes (37C) in the (CellBind) incubator
Centrifuge for 10 minutes in the CellBind
centrifuge (programmed protocol, 3 phases)
Read the reactions
13
Test procedure
14
Centrifugation steps

• 1. 2 min. 75 x g cells are separated from the
  serum in the incubation compartment
• 2. 1 min. 200 x g cells are spun down on the
  top of the gel matrix, where the sensitized cells
  bind to the active gel matrix
• 3. 7 min.1790 x g cells that are not
  sensitized are spun down through the gel to the
  bottom

15
Reaction strength, interpretation
4 reaction All sensitized cells will adhere on
the top of the gel matrix 3, 2, 1 and
() Cells at the top and at the bottom -
reaction Non-sensitized cells will form a
discrete button at the bottom
16
Application of CellBind
Detection and identification of red cell
antibodies Cross-matching Modified direct
anti-globulin (affinity) test (DAT) Typing
17
CellBind products

• CellBind Screen, 48 cards
• CellBind LISS, 250 mL 100 mL
• CellBind Direct, 96x IgG,C3d,Ctl
• CellBind P2, 2x10 mL
• CellBind P3, 3x10 mL
• CellBind ID16, 16x3 mL