High-Throughput Mannose Glycans Screening of Crude and Purified mAb and Non-mAb Protein Sample – Sartorius Octet® GlyM Kit

High-Throughput Mannose Glycans Screening of Crude and Purified mAb and Non-mAb Protein Sample – Sartorius Octet® GlyM Kit


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  • Source: SARTORIUS

  • Date: 11 Oct,2021

Glycosylation, an important post-translational modification, is a key critical quality attribute (CQA) that influences product safety and efficacy and should be monitored when developing new biological drugs. Among various glycan structures, mannose is especially important as it affects a protein’s plasma clearance and reduces its in vivo half-life, negatively impacting drug efficacy.
While process conditions, media and feed formulations are critical factors in the optimization of protein mannosylation, optimal cell line selection can be a limiting step; early screening for mannose content can facilitate the selection of clones that produce proteins with favorable glycan profiles during cell line development (CLD).

The Octet® GlyM Kit enables high-throughput mannose content screening in crude cell culture samples during early clone selection stages in the CLD process. When combined with titer analysis, it expedites the selection of the top performing cell lines producing proteins with the most favorable glycan (mannose) profile (Figure 1).

Only clones which are the most suitable will move downstream for further optimization.

Reliable data at this early stage supports shorter lead times and prevent later failures. Assay Workflow the GlyM Kit utilizes mannose specific lectin which is immobilized onto the biosensor surface (Figure 2). This surface binds to mannose structures in the sample.

The secondary amplification steps increase the binding signal from protein of interest (POI) and negates the minimal signal from host cell proteins (HCPs). This increases assay sensitivity, facilitating the analysis of crude cell culture samples without the need for purification or enzymatic digestion. This essentially eliminates the need for any sample preparation beyond a dilution step, simplifying the mannose screening workflow for users. Additionally, the GlyM Kit reduces time to results and increases sample analysis throughput. As a result, 96 clones can be screened in under 1 hour on the Octet® RH96 system.

The GlyM Kit comes with the biosensors and reagents required to screen mannose content in up to 96 samples and is compatible with Octet® systems with at least 4 channels. Relative Mannose Content Screening the GlyM Kit enables the screening and ranking of crude samples based on the relative mannose content in the POI. The kit provides ranking results for samples with mannose content that differs by 4% or more (Figure 3, see samples with 3% vs. 8% and 10% vs. 14% Mannose content). Users can assign all screened clone samples into groups depending on the mannose content (e.g., high, medium or low) and focus further development effort on the desired group of clones only. Data Analysis Octet® Analysis Studio software combines titer data with mannose content data for more in-depth analysis that facilitates selection of the best clones.

Source: Octet RED384 System (sartorius.com) Mannose Glycans Content Screening for Human-mAb Samples Using the Octet® GlyM Kit

The Octet® GlyM Kit is designed to provide cell line development and upstream bioprocessing groups with a ready to use kit for the rapid relative mannose content screening of both purified and cell harvest antibody samples. Unlike traditional techniques, the Octet® platform can also monitor other functional critical quality attributes (CQAs).

The ability to view and choose from desired titer and mannose levels at the same time provides more in-depth knowledge that facilitates more informed decisions in the early selection of optimal clones during cell line development.In this application note, a novel approach for relative mannose content screening and ranking of human monoclonal antibody (mAb) samples is showcased; using a well characterized mAb from the National Institute of Standards and Technology (NIST) as well as samples developed at Novartis.

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