The formulation of biological antibodies is the foundation for ensuring the production, storage, and transportation of stable products. The development process of formulas requires the use of high-throughput methods to reliably and quickly screen a large number of formula conditions, in order to determine the optimal formulation and "good conditions". A widely used method is to conduct thermal denaturation testing and derive relevant parameters, such as Tm and Tonset, to perform graded screening of formula conditions and the stability effect of excipients.

The traditional label free thermal denaturation technology is limited by flux, such as sample concentration, volume, consumable cost, and measurement speed, which makes the formulation end selection process with hundreds or thousands of potential conditions a challenge.

In this report, we will demonstrate the use of SUPR-DSF instrument to analyze and select the formulation of trastuzumab under 96 different conditions based on thermal denaturation parameter. When screening stabilizers, the results obtained are consistent with those obtained by differential scanning calorimetry: DSC method. This formulation has also been commercialized for the targeting of Herceptin. The screening of the formula was carried out in a 384 well microplate, and all results were obtained in less than 1.5 hours. This screening of sound flux can be integrated into laboratory automation systems, enabling the selection of thousands of samples per day.

Result:

The measured denaturation curves (Figure 1 and Figure 2 are examples) show two independent denaturation processes. The Tm value, van der Waals enthalpy (Δ H), and Tonset value were determined by performing * small two multiplication fitting on the denatured data using the standard three state model [1], and are listed in Table 1.

The table also lists the excipients screened to improve antibody stability. The experimental results obtained based on the three state model are consistent with the DSC data of trastuzumab [2], both reporting two Tmn values near 700C and 82oC. Considering the changes in the sample and solvent, the Tm value is consistent with the reported value [2].

In addition to similar Tm values, SUPR-DSF data confirms that histidine and trehalose dihydrate neither hinder nor enhance the conformational stability of trastuzumab, which are all components of the formulation of trastuzumab containing berberine [3]. These two points help demonstrate the consistency of SUPR-DSF measurements and the ability of high-throughput testing.