Screening and Production of Recombinant Human Proteins: Protein Production in Insect Cells

Mahajan, P.; Strain-Damerell, Claire; Mukhopadhyay, S.M.M.; Fernández-Cid, Alejandra; Gileadi, O.; Burgess-Brown, N.

doi:10.1007/978-1-0716-0892-0_5

Cited by 4 publications

(1 citation statement)

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“…Yeast are cheaper, easier to genetically manipulate and grow than other eukaryotic hosts, are capable of some essential PTMs [ 2 ], but lack essential lipids, impairing human MP production [ 33 ]. Insect cells present an alternative to yeast [ 27 , 28 ] but use baculoviral vectors for construct delivery which is time consuming [ 28 ]. We find that the ExpiSf system [ 34 ] improves production efficiency by maintaining higher cell densities.…”

Section: Choosing An Expression Systemmentioning

confidence: 99%

The high-throughput production of membrane proteins

Birch

Quigley

2021

Emerging Topics in Life Sciences

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Membrane proteins, found at the junctions between the outside world and the inner workings of the cell, play important roles in human disease and are used as biosensors. More than half of all therapeutics directly affect membrane protein function while nanopores enable DNA sequencing. The structural and functional characterisation of membrane proteins is therefore crucial. However, low levels of naturally abundant protein and the hydrophobic nature of membrane proteins makes production difficult. To maximise success, high-throughput strategies were developed that rely upon simple screens to identify successful constructs and rapidly exclude those unlikely to work. Parameters that affect production such as expression host, membrane protein origin, expression vector, fusion-tags, encapsulation reagent and solvent composition are screened in parallel. In this way, constructs with divergent requirements can be produced for a variety of structural applications. As structural techniques advance, sample requirements will change. Single-particle cryo-electron microscopy requires less protein than crystallography and as cryo-electron tomography and time-resolved serial crystallography are developed new sample production requirements will evolve. Here we discuss different methods used for the high-throughput production of membrane proteins for structural biology.

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Section: Choosing An Expression Systemmentioning

confidence: 99%

The high-throughput production of membrane proteins

Birch

Quigley

2021

Emerging Topics in Life Sciences

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Glycosylation in Drosophila S2 cells

Xu,

Tong,

Zhang

et al. 2024

Biotech & Bioengineering

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In recent years, there has been a remarkable surge in the approval of therapeutic protein drugs, particularly recombinant glycoproteins. Drosophila melanogaster S2 cells have become an appealing platform for the production of recombinant proteins due to their simplicity and low cost in cell culture. However, a significant limitation associated with using the S2 cell expression system is its propensity to introduce simple paucimannosidic glycosylation structures, which differs from that in the mammalian expression system. It is well established that the glycosylation patterns of glycoproteins have a profound impact on the physicochemical properties, bioactivity, and immunogenicity. Therefore, understanding the mechanisms behind these glycosylation modifications and implementing measures to address it has become a subject of considerable interest. This review aims to comprehensively summarize recent advancements in glycosylation modification in S2 cells, with a particular focus on comparing the glycosylation patterns among S2, other insect cells, and mammalian cells, as well as developing strategies for altering the glycosylation patterns of recombinant glycoproteins.

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