Efficient production of an antibody Fab fragment using the baculovirus–insect cell system

Lepidopteran cells, an alternative for the production of recombinant antibodies?

“…Although there are exceptions, High Five cells (derived from the lepidopteran T ni) are generally more efficient at expressing and secreting recombinant proteins than Sf 9. 18,19 However, it should be noted that the N-glycosylation performed by High Five cells is not identical to general assumption, the transcriptional patterns of the P10 and PH genes are similar but not identical. 16 P10 gene is expressed a few hours before polyhedrin, thus leading to the overexpression of the H chains before the light chains and triggering of secretion of H chains dimers (H 2 ).…”

“…Although the exchange of the signal peptide sequence can significantly increase the production of some proteins, no significant enhancement was observed in immunoglobulin production after exchanging the signal peptide with the honeybee melittin, AcMNPV gp64 or Drosophila BiP sequences. 18 From the very first reports of Ig production in insect cells, the examination of whole cell lysates revealed the presence not only of fully functional H2L2 molecules, but also of unprocessed Ig polypeptides (e.g., precursor of light chains) or aggregates. 2 This indicated that the folding and assembly machineries may be unable to process all Ig chains synthesized during infection.…”

“…18,40,[43][44][45][46] It is very easy and fast to generate recombinant viruses expressing H or L bearing specific mutations and to determine the impact of these mutations on the specificity or activity of a given antibody. 21,22,41 This strategy has been successfully used to analyze the catalytic activity of the 6D9 monoclonal antibody and the specificity of the PAC1 antibody that binds to the integrin αII b β3 on activated platelets.…”

“…17 Aggregation may in turn also explain the low expression rate observed in these conditions. 18 In conclusion, to obtain adequate and correctly folded Igs in insect cells, it is important to use promoters that direct the and J chain. After binding to the poly-immunoglobulin receptor (pIgR) dimeric IgA is transported across the epithelial barrier to be secreted on the luminal side of the mucosa.…”

“…46 Beside these full-length immunoglobulins, many different antibody-derived molecules have been produced e.g., mono-specific single-chain fragments, scFv, [47][48][49][50] bi-specific scFv, 51 Fab. 18,[52][53][54] The expression of scFv is more reliable compared with that of the E. coli system and does not require any re-folding step. Fusion proteins such as chimeric-hormone-antibody molecules that achieved in human cells and may even be immunogenic (see below, "Glycosylation in insect cells").…”