New Technology Opportunity

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Who We Are

Bondwell Technologies was founded to commercialize a revolutionary biomaterial-based platform technology with the unique ability to safely immobilize and stabilize high concentrations of protein in an inexpensive and facile process. Proteins are nature’s workhorse and provide structure, function, and regulation for almost every biological task. Biological processes, and therefore proteins, have been used in a broad range of industrial applications for centuries; however, isolating a specific protein for a specific use is highly beneficial for high-impact markets such as energy and medicine. Unfortunately, isolating protein is notoriously difficult and costly because proteins are easily degraded and difficult to store and produce. Bondwell Technologies’ platform solves these problems by inexpensively producing proteins bound to a novel material that protects them from degradation. Our versatile platform technology can be used to develop a portfolio of products in multiple fields including energy, environment, and medicine.

Technology Summary

Proteins have amazing functions that can be harnessed for many applications in biotechnology. However, proteins are fragile, largely limiting their practical use. Proteins are difficult to incorporate into materials without either harming the protein’s activity or making the materials toxic. Further-more, proteins rapidly degrade and inactivate once incorporated into materials. Bondwell Technologies is uniquely able to make new materials from an engineered protein. These materials can be in the form of a film, a fiber, annealed into a mesh or electrospun as a membrane (Figure 1).

Most importantly, only our material can incorporate active large, complex proteins via protein fusion, eliminating the need for chemical crosslinkers. In the protein fusion approach, the gene encoding a desired protein is placed adjacent to the gene for our material protein. In bacteria or yeast, this gene fusion produces the protein fusion as a single polypeptide. After the distinct segments of the fusion protein independently fold, the material portion of the fusion protein drives materials formation.

Bondwell BioPure Performance Data

We have been evaluating our Protein A chromatography membranes in-house for antibody capture in flow systems analogous to commercial membrane systems already on the market such as those offered by GORE, Sartorius, and Cytiva. Our current membranes are being fabricated through an electrospinning process onto a thin substrate, followed by cutting and placement into a capsule for analysis. Both elements – the capsule and the membrane – play an interconnected role in performance. As we continue refining our production process, some natural variability exists in product metrics due to our current scale.

Project Overview

Bondwell Technologies has developed a membrane-based affinity chromatography device. Currently, our device is optimized for performance at the 0.1 mL scale. The purpose of the evaluation would be to:

Carry out testing in a way that is mutually beneficial to both Bondwell and the test site
Validate Bondwell’s devices by an external lab for feedback and improvements

Resource Requirement

mAB targets

100mg (1-5mg/mL) Pure (post-ProA) target for DBC experiements
100mg per 100 cycles of each target as Clarified Cell Harvest

Buffers

10L Equilibration buffer per 100 cycles
2.5L Wash buffer per 100 cycles
1L Elution buffer per 100 cycles
1L CIP (0.1M NaOH) per 100 cycles if required

Ideally use of an AKTA Avant/Pure 150mL/min Systems

Analytical support e.g. HCP, product concentration, ProA Leaching ELISA (other to be confirmed)

Proposed Studies

Study 1: Productivity Comparison

To evaluate the productivity range of our membrane capsules, analysis could be performed at different residence times (e.g., 1 s, 3 s, 6 s, 15 s, 30 s) and loading feeds. We estimate this would take ~3 hrs at the five example residence times.

Estimated resource requirements (per capsule):

Equilibration buffer (e.g., PBS): 25 mL
mAb loading: 5 mg per sun, 25 mg total
Wash buffer (e.g., PBS): 25 mL
Elution buffer (e.g., 0.1 M glycine, pH = 2.5): 10 mL
Regeneration buffer (10 mM NOH, every 5 cycles): 7 mL

Analytical:

HCP Reduction: CHO HCP ELISA
mAb quantification during loading and elution: ELISA or HPLC
Protein A quantification in the eluate: Protein A ELISA

Study 2: Cyclability, 20 cycles

The high flow rates capable of Bondwell’s capsules means that our membranes can be run with relatively short cycle times. We have been using a cycle time of ~15 minutes (residence time of 6 s) specifically for our current prototype capsule. Given this, we could run 20 cycles in a timespan of ~5 hrs to demonstrate the reusability of our system. Our proposed resource requirements are below and can be changed dependent on client desire, for example, running additional cycles.