1. Introduction to the Project

1.1 Introduction to technical achievements

This achievement uses a sensing chip based on porous anodized aluminum oxide (PAA), which can quickly detect single base mutations in tumors through non amplification technology. After multi-step modification, PAA bound to streptavidin (SA) is obtained, which is then combined with biotinylated dCas9 protein (Biotin dCas9) based on CRISPR/Cas9 system, sgRNA specifically bound to the target sequence, and the target sequence to be tested. Then, a CRISPR-PAA electrochemical nucleic acid detection platform is constructed by using the principle of electrochemical ion current rectification (ICR). This platform is used for gene detection of single base mutations in diffuse large B-cell lymphoma (DLBCL), breast cancer, laryngeal cancer, snoring and other diseases. By now, through academic cooperation and after ethical approval is obtained, more than 400 clinical samples including diffuse large B lymphoma and breast cancer have been detected, which is consistent with the sequencing results.

1.2 Technical analysis and advantages

1.2.1 Core technical analysis

Professor Liang Chongyang, the technical leader of the project, has been committed to the research and production of “gene editing combined with electrochemical non amplification gene mutation detection reagents and instruments”, which is a pioneering product in China and also in the world. Professor Liang Chongyang has mastered the core technology of gene editing combined with electrochemical non-amplification gene mutation detection, and is at the world’s advanced level in the development of products and applications related to gene editing combined with electrochemical non-amplification gene mutation detection.

1.2.2 Technological innovation and advantages

This project aims to meet the huge sequencing market demand brought about by precision medicine, and has developed a non-amplification genomic single base mutation rapid detection chip and supporting instruments. Compared with traditional sequencing techniques, the detection time is shorter, completed within 1 hour, and the sequencing method takes 15-20 days; without the need for PCR amplification purification laboratories and professional bioinformatics staff, industry professionals can also conduct testing in ordinary rooms.

The core patent has been authorized; the production process and quality research of the reagent small-scale trial have been completed; the principle engineering machine has been developed and completed; through academic cooperation, after ethical approval is obtained, more than 400 clinical samples including large B lymphoma and breast cancer have been detected, which is consistent with the sequencing results.

1.3 Main products and applications of technical achievements

Compared with traditional sequencing techniques, the CRISPR-PAA electrochemical nucleic acid detection technology has the technical advantage of short detection time and is not limited by the testing room. In the past, patients needed to go to the provincial capital city for testing, and this technology can also be carried out in local or county hospitals, which is in line with the current trend of digital diagnosis and treatment and improves the accessibility of medical care for the public.

1.4 Progress stage of technological achievement commercialization

By now, the fermentation and purification process of dCas9 protein has completed small-scale and pilot tests, and is ready for engineering.

Through multiple explorations and verifications of modification conditions, it has been possible to mass produce AAO membranes modified with streptavidin.

By optimizing the conditions such as reagent composition and freeze-drying curve, the required reagents for detection are made into freeze-dried balls for storage and transportation. Small and medium-sized tests have been completed, and the freeze-dried products can be stored for more than 2 years.

The development of the prototype is completed, and the instrument is compact and lightweight, suitable for POCT detection.

1.5 Technical team

Liang Chongyang, male, 42 years old, is a professor and doctoral supervisor at the School of Pharmacy of Jilin University. He is also the Director of the Modern Biology Teaching and Research Office at the School of Pharmacy of Jilin University and a part-time scientific consultant at Changchun Baike Biotechnology Co., Ltd. The research and development directions include recombinant protein drugs, vaccines, high-throughput targeted drug screening technology, single-cell sequencing, and spatial transcriptomics technology. The team under his leadership has completed preclinical development research for multiple new drugs and has been approved by the US FDA and China NMPA for clinical trials; he has developed new single-cell sequencing and spatial transcriptome detection methods using the supramolecular assembly technology. He has published 60 papers in academic journals such as Analytical Chemistry, Lab on a Chip, ACS Sensors, Nanoscale, Biosensors & Bioelectronics, and led more than ten projects including the National Natural Science Foundation of China, National Key Science and Technology Equipment Project, and Jilin Provincial Department of Science and Technology Project. He has established Shanghai Pumafu Pharmaceutical Research and Development Co., Ltd.

Li Hongrui, the legal representative of the company, male, graduated from Jilin University in 2011 with a doctoral degree. After graduation, he stayed on campus to teach at the School of Pharmacy of Jilin University, and was responsible for laboratory management and research work at the Key Laboratory of Yeast Expression Recombinant Protein Drug Development at Jilin University. He has been engaged in research on the anti-tumor mechanism of fungal immune regulatory protein rLZ-8, and participated in the industry university research transformation of the project results. He has cooperated with enterprises to build factories and constructed GLP laboratories for the project. The project results have been approved for clinical trials of new drugs by China’s NMPA and the US FDA. His main research direction is the development of recombinant protein quality research technology, and he focuses on using ultra-high resolution microscopy imaging systems to study cell transport, intracellular ultrastructure analysis, and related basic research. He has participated in 3 projects funded by the National Natural Science Foundation of China and 5 projects funded by the Provincial Department of Science and Technology. He has published 12 academic articles, including 7 indexed by SCI. He will participate full-time in the establishment and operation of Changchun Pumafu through the method of leaving the job to start a business.

2. Market Potential Analysis

Tumor molecular diagnosis, as a sub field of molecular diagnosis, has developed rapidly under the influence of national policies and market demand growth. Due to the late start of related development, the market size of China’s tumor molecular diagnosis and testing industry is still relatively small compared to its potential market total. The tumor molecular diagnosis and detection market includes PCR based detection and NGS based detection. The main competitors in the industry are also based on PCR detection and NGS detection products, and there is no CRISPR-PAA detection platform like the company’s CRISPR/Cas system based non-amplification detection products. There is currently no medical device enterprise in China that uses the non-amplification nucleic acid testing technology.

Tumor molecular diagnosis, as a sub field of molecular diagnosis, has developed rapidly under the influence of national policies and market demand growth. Due to the late start of related development, the market size of China’s tumor molecular diagnosis and testing industry is still relatively small compared to its potential market total. It is expected that by 2030, the market size of the tumor molecular diagnosis and detection industry will have been 72.5 billion yuan, and the detection scale based on NGS will be 49.1 billion yuan; the scale of tumor molecular diagnosis and detection based on PCR is about 23.4 billion yuan, both of which are amplification detection technologies.

The tumor detection market in our country has enormous potential, with an annual medical expenditure of over 220 billion yuan for malignant tumors, which has become an important expenditure for households and medical insurance funds.

3. Financing needs and financing purposes

The project is planned to raise 1 million yuan, mainly for product research and marketing. The project is expected to have completed the engineering acceptance of the clean workshop by the end of 2025, and apply for clinical trials of non-amplifying gene mutation detection kits in October, 2026.

4. Analysis of Technical Achievements and Benefits

4.1 Economic benefits

After the achievement is transformed and launched, the product will gradually occupy the market share of diffuse large B lymphoma testing, from 5% in the first year to 20% in the third year, and the estimated output value will reach 15 million yuan in the third year.

4.2 Social benefits

The project will establish the first non-amplified gene mutation detection platform in Changchun, which will further cover the detection kits for guiding drug use in gene mutation detection of lung cancer, breast cancer, hypopharyngeal cancer, colorectal cancer and other cancers, and cooperate with the country to promote the accessibility of medical resources so that patients can carry out gene mutation detection in prefecture level cities or county hospitals. It is expected that within five years, with the posture of a unicorn enterprise, this technology platform will leverage the billions of medical diagnosis markets. Combined with digital medicine and medical consortia, it will undoubtedly make important contributions to improving public health.

5. Contact Method

Contact person: Liang Chongyang

Tel: 17790098441