Afuco™ Platform
company’s services is the Afuco™ platform, used for the generation of ADCC enhanced antibodies by means of transient expression in CHO cells. By using homologous recombination technology, we obtained a fucosyltransferse-engineered stable CHO cell line, which produce defucosylated monoclonal antibodies with enhanced affinity for FcγRIIIa and enhanced ADCC activity.
2. Our Afuco™ platform 2.1 Fut8-/- CHO-K1 Stable Cell Line Establishment 2.1.1 Project Overview Objective Establish the Fut8 knockout cell line using CHO-K1-GS-/- cells. Description 1)
Design gRNA according to the Fut8 sequence (Gene ID: 100751648).
2)
Construct the knockout vector.
3)
Transfect CHO-K1-GS-/- cells
4)
Screen for positive cell clones.
Target Gene Fut8 (ID: 100751648) Plasmid
pGK1.1 (Amp) Host Cells
CHO-K1-GS-/- cells Final Delivered Items
Item
Description
Size
Storage
1
Fut8 knockout (-/-) cells
1 vial
Liquid nitrogen
2.1.2 Methods & Results Part I Preliminary test 1)
Optimize electroporation condition The electroporation condition has been optimized.
2)
Test the minimal absolute lethal dose of Puro The minimal absolute lethal dose of Puro has been tested.
3)
Test the clonability of CHO-K1-GS-/- cells The clonability of CHO-K1-GS-/- cells has been tested.
Part II Design gRNA & construct knockout vector 1)
Design gRNA Gene Name: Fut8 Species: Chinese Hamster mRNA Refseq: XM_003501735 Sequences: TTTGGTCTCAAAAACCAAAAAACTTAAATCTGTTGATTCCAGGTTCCCATATATTCTTGGATATGCCAATTACTTTTTCTGTAAGCAAG TGTTTCATAAAACTTTTACTTAACTTTCATATTGACCTGTACTATTCAACATTCAGCTATGTTAAAGTATTTGTGAAGTGTTTTGAAATG ATTTTATATTTTCTAAGGTGAGAATAAATGAGAAAATGTTTTAATATGTCTCCAGTGCCCCCATGACTAGGGATACTAATTGAGTACCA
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