4 December 2018
AbTheneum is SCT’s patented process for antibody discovery. Antibodies are analyzed phenotypically and genotypically from individual antibody secreting cells, examining thousands of cells simultaneously. Antibodies are rapidly tested for binding activity and specificity by screening against multiple antigens of interest. Messenger RNA encoding the cognate VH and VL are analyzed by Next Generation Sequencing (NGS) technology and correctly paired.
The process starts with the SCT picoliter device, made of polydimethylsiloxane (PDMS, an elastomeric material). One picoliter device contains approximately 90,000 microwells to trap single antibody-producing cells. Each microwell in the picoliter device can be imagined as one well on a 96-well plate—only with 1,000,000-times smaller volume! That means 1 SCT picoliter device is equivalent to almost 1,000 96-well plates.
This small volume allows fast kinetics. A single plasma cell is a professional at antibody secretion, pumping out antibodies at rates as high as 2,000 molecules per second. In a tiny SCT microwell, the antibody concentration goes up fast.
Antibody-secreting cells are deposited into the microwells. The secreted antibodies are captured by pressing the picoliter device against a capture slide. While pressed against the SCT picoliter device, the capture slide captures antibodies from each well containing an antibody-secreting cell. Once the picoliter device and the capture slide are separated, the capture slide can be assessed off-line, or separate from the cells. This antibody array means thousands of different antibodies can be assessed for their binding properties at the same time at low reagent cost.
The binding activity of antibodies captured onto the antibody array is determined by flowing fluorescently labeled screening molecules over the surface. A microarray scanner images the array for spots or “hits” against the antigen (green) or all other IgGs (red).
The antibody-secreting cells sitting in the SCT picoliter device are lysed and the encoding messenger RNA for the antibodies are captured. An addressable DNA microarray is pressed against the picoliter device to capture mRNA from each individual cell. The mRNA is converted to cDNA and sequenced by NGS. The DNA sequences can be correlated with the antibody binding data gathered from the antibody array using a sequence barcode embedded in the DNA microarray.
The results from AbTheneum are a massive library of information on the immune response. We call these results the Antibody Atlas. The sequences of VH and VL correlated with binding properties of the antibodies are all clustered into related families. The members of the family all likely bind to the same epitope, or region, of the antigen.
SCT’s AbTheneum platform is unparalleled in the industry. If you are struggling to find those long sought-after antibodies, you will want to try AbTheneum.
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