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Stability Analysis

Creative Biolabs is specialized in providing bispecific antibody (BsAb) analysis services, and now offers customers the elaborate analysis strategy for completely assessing your BsAb stability. Based on our advanced instrument and extensive expertise, we can provide various analytical approaches, including Thermal Stability Measurement, Aggregation Measurement, and In vitro Serum Stability Measurement.

Aggregation Measurement

The use of antibodies as biotherapeutics has notably increased in recent years. During the production process, one primary concerns is the propensity of a biotherapeutic antibody to aggregate. Protein aggregation has crucial effects on drug potency and results in highly undesirable immunological effects. Therefore, it is important to measure aggregation conditions of therapeutic antibodies. Creative Biolabs provides various techniques to measure aggregation of BsAbs or other therapeutic antibodies, such as Dynamic Light Scattering (DLS), Electrospray Differential Mobility Analysis (ES-DMA), Sedimentation Velocity Analytical Ultracentrifugation (SV-AUC), and Field Flow Fractionation (FFF), etc.

Stability Analysis

Thermal Stability Measurement

Among many factors affecting proteins aggregation, temperature is an important environmental factor. Temperatures of 40 to 70 °C usually lead to the loss of protein activity, on account of protein unfolding and aggregation. Since the probability of protein denaturation increases with higher temperature, heat resistance is indicative of long-term stability at storage temperatures. Thus, heat is typically used as a source of stress for measuring the thermal stability of protein. Creative Biolabs provides several techniques to measure thermal stability of BsAbs or other therapeutic molecules, including Differential Scanning Calorimetry (DSC) and Differential Scanning Fluorimetry (DSF). DSC is a thermal analysis apparatus for determining how physical properties of a sample change, with the variation of temperature against time. DSF makes use of the high thermal control, sensitivity, and expanded availability of RT-PCR machines to determine protein thermal stability.

In Vitro Serum Stability Measurement

Measurement of the in vitro serum stability is important given that rapid degradation of novel therapeutic BsAbs in plasma may occur, which generally leads to poor in vivo efficacy of the molecule and even toxicity. Stability in plasma is also a factor that may affect the accuracy of in vitro and in vivo data, for example, the plasma protein binding data. Plasma stability test is a useful strategy for screening drugs that require rapid conversion in plasma. Creative Biolabs provides various techniques to measure thermal stability of BsAbs or other therapeutic molecules, such as Enzyme-linked Immunosorbent Assay (ELISA) and Flow-cytometry Assay.

Stability Analysis

Figure 2. Immunoreactivity and avidity of BsAb. (A) and (B) show the flow cytometric results of binding of bsAb FabCD19xCD5 and its parental antibodies to lymphoma lines Raji and Jurkat. (C) and (D) show the results of flow cytometric binding competition assay comparing the binding strength of bsAb versus the parental mAb (Lüttgau, S. 2013).

With our well-established BsAb analysis platform, the experienced scientists at Creative Biolabs are dedicated to helping you develop BsAbs using a full range of techniques. Our high-quality stability analysis services will greatly contribute to the successes of your projects. Creative Biolabs also provides various other services regarding BsAb development. Please feel free to contact us for more information and a detailed quote.

References

1. Lüttgau, S.; et al. Immunotherapy of B-Cell Lymphoma with an Engineered Bispecific Antibody Targeting CD19 and CD5. Antibodies. 2013, 2(2): 338-352.
2. Spink, C. H. Differential scanning calorimetry. Methods in cell biology. 2008, 84: 115-141.

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