Technology

Technology Overview
Technology for Screening and Production

Technology Overview

ApoLife has developed proprietary expression technologies which include expression vectors, defined media formulations free of animal derived components, high density fed batch fermentation processes, and yeast host strains developed for high levels of secretion of recombinant proteins and antibodies.

The advantage of ApoLife’s proprietary “Twin Cassette®” plasmid is that it favors equimolar expression of antibody chains from a single plasmid, resulting in a greater yield of functional antibodies.

Using proprietary yeast strains, ApoLife has demonstrated successful expression of full-length functional monoclonal antibodies and antibody fragments including fusion proteins, which are secreted into media. Preliminary glycoanalysis of antibodies produced from ApoLife’s proprietary yeast strains do not show hyper mannosylation.

Advantages of S. cerevisiae expression system

  • Secreted expression of full length antibodies
  • No methanol handling required for fermentation
  • No antibiotic selection of plasmids
  • Product safety-no risks from endotoxins, animal viruses, prions or oncoproteins
  • Reduced product development time to the market
  • Genome of S. cerevisiae completely sequenced and extensively characterized enabling the easy manipulation of host yeast strains
  • Production process transferable to current microbial manufacturing facilities
  Features Advantages
1 Twin Cassette® Plasmid Equimolar Expression of Both Antibody Chains
2 High Copy, Highly Stable Plasmid High expression of antibodies and proteins
3 Proprietary Yeast Strains Selected for High Expression & Secretion
4 Rapid Cell Growth (90 min/generation) Shorter Production Time
5 Nutritional Markers for Selection Less Regulatory Hurdles
6 Secretion of Antibodies into Media Lower Purification Costs and Rapid process development
7 Rapid Growth to High Density using inexpensive, defined media Lower cost of production
8 Fed Batch Fermentation 7 days vs. 21 days in Mammalian Cells Lower infrastructure cost
9 Scalable Rapidly Scalable from 1 L to
50,000 L


ApoLife’s Twin Cassette® Plasmid



Production strain development timeline


Technology for Screening and Production

Cost-effective Antibody Screening and Production

Accelerated Selection of Target Antibodies

ApoLife’s unique platform enables rapid screening of target antibody candidates using its proprietary Plug-N-Play® Twin Cassette® vectors.

Several companies use hybridoma or phage display generated clones as starting material for selecting target specific recombinant antibodies. Currently companies screen several mAbs before bringing one candidate to preclinical development using HEK 293 transient cells or CHO cells. ApoLife's robust yeast system provides milligram quantities of target antibodies (as shown in the following table) in the shortest time compared to the current systems.

With this capability, companies can test a greater number of therapeutic candidates thus increasing the probability of finding a suitable candidate.

Rapid Identification of Target Antibody Clones

Method 2 mg IgG (days) 100 mg IgG (days)
ApoLife Yeast 3-5 days/100 ml 5 days/2 L
HEK 293- Transient 10 days Impractical
CHO- Stable Line 30-60 days 3-6 months