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.

Product Overview 

LAMP lyophilized reagents refer to the lyophilization treatment of various components required for LAMP reactions (including Bst enzyme, reverse transcriptase, dNTPs, MgCl₂, buffer, and primers, etc. ), preparing them into stable powder or lyophilized ball products. After reconstitution with water, LAMP amplification reactions can be carried out directly. Compared with traditional liquid LAMP reagents, lyophilized reagents have strong stability, are easy to use, and can be stored at room temperature, greatly reducing the cost of cold chain transportation.

Key Features

1.   Cear, pink-to-yellow visible detection of amplification.

2.   Fast, readout of positive amplification analyzed by eye in 20 mins at 50 copies/T.

3.   Good thermal stability after 37 C for 7 days.

Performance Data

Hzymes provides LAMP/RT-LAMP lyophilized premix and products, and also provides customized CRO/CDMO lyophilization services, meeting the diverse needs of customers and customizing special lyophilized products for customers.

Transportation and storage conditions

Transported with Dry ice, stored at -25 ～ - 15℃. Avoid repeated freezing and thawing, the product is valid for 12 months.