Lytic Solutions LLC provide high-quality, cost-competitive custom mammalian cell protein-expression services. We have significant experience and expertise in the production of Fc- and His-tagged fusion proteins, in addition to, scFvs, recombinant monoclonal antibodies, and diabodies via transient transfection of suspension-cultured mammalian cells. We provide numerous options to best suit your needs from small-scale expression screening (50 µg-1 mg protein produced) to larger scale production (10 mg to grams of protein produced). We have culture volume capacities of 30 mL up to 50 L for single vessel runs, so we can accommodate a variety of protein expression runs via transient-transfections or culturing of stably transformed cell lines.   Contact us to find out how Lytic Solutions can help you with your mammalian cell protein expression needs.

General Information:

Suspension-cultured cell lines typically used:

Chinese hamster ovary (CHO) cells

Human embryonic kidney (HEK) cells

Expression vector and protein format options:

(1)  Lytic Solutions will use a provided expression vector or we will custom construct one using Lytic Solution vector backbones.

a.     Lytic Solutions have several high copy-number, strong constitutive promoter vectors available for expression use.

(2)  The expressed protein can be designed to be secreted or retained intracellularly.

(3)  Epitope-tagged or tag-free

a.     Epitope-tags used for purification: poly-histidine (His6-His10), Fc-fusion

b.     Engineered with/without Tobacco etch virus (TEV) protease cleavage site for tag removal

(4)  We are willing to work with you to provide alternative options that meet your needs.

Typical workflow for transient-transfection custom production:

(1)  Provide Lytic Solutions with the amino acid sequence for expression

(2)  Codon-optimized coding sequence synthesis

(3)  Cloning CDS into vector of choice

(4)  Prepare bulk, transfection-grade plasmid DNA using Lytic Solutions in-house methods.

(5)  Transient transfection and suspension-culturing of CHO cells

(6)  Purification of produced protein

a.     We use high-quality, pristine Lytic Solutions IMAC or Protein A-agarose resins for all affinity purifications.

b.     For untagged proteins, conventional chromatography procedures with high-quality, pristine resins will be developed and used.

(7)  Protein buffer-exchanged into agreed-upon final buffer

a.     Final product can be supplemented with preservatives or sterile filtered to minimize degradation and improve stability.

(8)  Typical protein analyses performed:

a.     UV-Vis spectroscopy

b.     Protein quantification assay (i.e. BCA)

c.     SDS-PAGE

d.     HPLC with light absorbance detection (i.e size-exclusion chromatography (SEC) and ion-exchange chromatography) (Optional, available)

(9)  Protein is overnight-shipped cold to client. All produced proteins will include a product report that includes final protein specifications and analysis data.

Figure Legend:

(A)  Small-scale screen for expression of seven scFv-Fc fusions. (SDS-PAGE sample protein load based on volume)

(B)  Production of a functional diabody-His6. (SDS-PAGE sample protein load: 4 µg)

(C)  Production of a recombinant monoclonal antibody. SDS-PAGE analysis was performed under both reducing (+ DTT) and non-reducing (-DTT) conditions. (SDS-PAGE sample protein load: 7 µg)

(D)  Large-scale production of two secreted His-tagged fusions. (SDS-PAGE sample protein load: 3 µg)

(E)  Large-scale production of a Fc-fusion with analysis by reducing SDS-PAGE (left) and size-exclusion high performance liquid chromatography (SEC-HPLC) (right). (SDS-PAGE sample protein load: 3 µg)