Unlocking a New Era in PCR: Whole-block Taq Antibody Leads Efficient and Convenient Amplification

Background

The perfect combination of Taq DNA polymerase and PCR technology has made PCR a practical tool, widely used not only in scientific research but also in areas such as pathogen detection, genetic testing, tumor companion diagnostics, as well as in forensic science, environmental monitoring, and food safety. With the advancement of technology, PCR is evolving towards being more convenient, sensitive, and cost-effective.

To meet the demand for efficiency and convenience, pre-mixed reagents are increasingly replacing stepwise reagents as the mainstream option. During large-scale pandemic responses such as COVID-19 and African Swine Fever, testing organizations have placed more emphasis on the stability of pre-mixed reagents to ensure accurate test results.

The antibody-modified hot-start Taq DNA polymerase has been widely promoted in various practical applications due to its ability to partially address non-specific amplification in PCR. However, in conventional probe-based test reagents, the 5’→3′ exonuclease activity of Taq enzyme can degrade key materials such as primers and probes, reducing amplification efficiency and reagent stability. Market competition and globalization pressures have driven reagent manufacturers to further improve reagent stability, requiring pre-mixed reagents to maintain excellent performance after being stored under pressure at 4°C for 14 days or 37°C for 7 days.

Therefore, companies around the world have been researching more efficient Taq enzyme antibodies to completely block both 5’→3′ polymerase activity and 5’→3′ exonuclease activity of Taq enzyme. Since the outbreak of COVID-19, various manufacturers have introduced dual-function block antibody products, but none have been perfect. Either the blocking efficiency for both polymerase activity and exonuclease activity fails to exceed 90%, or the antibodies dissociate from Taq enzyme at temperatures above 55°C, leading to instability in pre-mixed reagents.

In order to discover the perfect Taq enzyme antibody and further improve PCR reagent performance, HZYMES used B-cell cloning technology to screen B cells enriched by magnetic bead sorting using the Beacon single-cell optofluidic system, and then PCR-amplified cDNA to directly construct linear expression frames for transient transfection, followed by ELISA screening and verification. After sequencing the selected antibody genes and verifying the binding strength and blocking functions, HZYMES’ research team found that different Taq enzyme antibodies exhibit a certain synergistic effect. It was generally believed that mixing two specific antibodies that individually block polymerase activity and exonuclease activity would perfectly block both functions of Taq enzyme, but the results were not ideal. However, by adding a third antibody to the mixture of two antibodies that moderately block 5’→3′ polymerase activity and 5’→3′ exonuclease activity, the dual-function blocking effect of Taq enzyme significantly improved.

Figure 1: Diagram of HZYMES’ Whole-block Taq antibody mechanism (gray represents 5’→3′ exonuclease activity domain, green represents 5’→3′ polymerase activity domain, and yellow represents 3’→5′ exonuclease activity domain).

The fully Whole-block Taq antibody product developed by HZYMES consists of an optimized combination of three antibodies (Figure 1). Compared to single or dual antibody mixtures available in the market, this product can block more than 99% of Taq enzyme’s 5’→3′ polymerase activity and 5’→3′ exonuclease activity at 70°C, offering superior blocking efficiency and stability. This ensures that non-specific amplification caused by mismatched templates or primer dimers is minimized, and degradation of primers or other key materials is avoided, thereby improving reagent stability. Additionally, it offers rapid enzyme activation, with over 95% of enzyme activity released within 30 seconds at 95°C.

(Participate at the end of the article, free trial)

Superior Product Performance

Whole-block:

blocksboth polymerase activity and 5’→3′ exonuclease activity with over 99% efficiency.

Wide Compatibility:

Works with various types of Taq DNA polymerase, including wild-type and mutant forms.

Fast Release:

More than 95% of enzyme activity is released within 30 seconds of heat activation at 95°C.

Strict Quality Control:

Ensures purity, concentration, and functionality, with rigorous testing for nuclease residue and host DNA contamination.

Quality Control Items	Criteria
Appearance	Clear, transparent liquid
Concentration	5±0.5 mg/ml
SDS-Purity	>95%
Endonuclease Residue	none
Exonuclease Residue	none
Nickase Residue	none
RNase Residue	none
Host Nucleic Acid Residue	none
Capping Efficiency	>99%
Activation Efficiency	>95%

Application Case Studies

Pre-mixed Reagent Stability:

Compared to similar products, HZYMES’ Whole-block Taq antibody offers higher stability at 37°C for 28 days, without a significant reduction in amplification efficiency.

Specificity in Multiplex PCR:

Reduces non-specific amplification products, as demonstrated in forensic testing (Figure 6).

Order Information:

In the future, Hzymes biotech will always remember its original intention and persist in meticulous cultivation in the IVD field. It will adhere to independent research and development, accelerate the construction of a world-class specialty enzyme production platform, and achieve import substitution of core enzyme raw materials in the field of biomedicine in China. It will collaborate with leading biopharmaceutical companies to expand their global presence and contribute to the advancement of the industry.