Background

The development of the lyophilization process is a systematic and complex process, involving multiple steps such as eutectic point testing, collapse temperature testing, sublimation rate testing, and lyophilization protectant screening. It is necessary to comprehensively analyze key influencing factors and conduct systematic experimental design to obtain a good lyophilization process.

Development process of freeze-dried products

In the development of a biochemical diagnostic enzyme raw material by Hzymes, due to the low collapse temperature of the solute and sample mixture and the low concentration of the mixture, the product after lyophilization appeared to be surface collapsed, non-uniformly dispersed, and unable to form a good lyophilized powder cake shape; more troublesome is that after lyophilization with only buffer components, the enzyme activity and the 37℃ heat stability incubation process of the lyophilized powder both decreased significantly (Formula 1.0).

The problem of product collapse is mainly due to the high primary drying temperature, so it can be solved by lowering the drying temperature. However, to form a honeycomb structure and maintain the stability of the enzyme, it is necessary to carry out the screening and combination of protectants.

In the preliminary screening, by evaluating their protective effects on enzyme activity, the advantageous lyophilization protectants were obtained: Formula 1.5 and Formula 1.6; further combination and concentration optimization of Formula 1.5 and 1.6 were carried out, and after 3 rounds of testing, the best formula with high lyophilization activity and good heat stability was finally obtained: Formula 3.4.

At the same time, through the optimization of primary drying temperature → protectant screening → lyophilization buffer solution formula, a good lyophilized appearance was finally obtained.

Figure note: The impact of lyophilization protectants on enzyme activity

Step 1: Due to the high primary drying temperature exceeding the collapse temperature, the phenomenon of product collapse occurred.

Step 2: Optimize the drying temperature to solve the collapse problem, but the product appears to be non-uniformly dispersed, mainly because the lyophilization structure is not rigid enough to form a good honey comb structure.

Step 3: Through the screening of protectants, the lyophilization morphology was improved, but due to the high surface tension between the sample and the vial, the product climbing phenomenon occurred.

Step 4: Adjust the lyophilization formula to improve the solution surface tension and obtain a good lyophilized product appearance.

Introduction

As a leader in the specialty enzyme industry, Hzymes has rich experience in the development and production of recombinant protein products, is good at E. coli and yeast expression systems, and has built a super clean enzyme purification platform and rich lyophilized formulation development technology through long-term project research and development, core technology accumulation, and industrialization in-depth cultivation. It can provide lyophilization platforms of different scales such as 0.4m², 0.5m², 2m², 10m², and 20m², which can match the production needs of small-scale trials, medium-scale trials, and industrialization.