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Hybridoma Technology Platform

Alpha Lifetech provides customers with quality assured antibody and recombinant antibody products and services.

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Hybridoma Technology Platform


Alpha Lifetech has rich experience in the field of antibody development. Over the years, Alpha Lifetech has established a comprehensive antibody development platform (including monoclonal antibody development, polyclonal antibody development, recombinant monoclonal antibody development, and other antibody preparation and development services). 

Alpha Lifetech provides customers with quality assured antibody and recombinant antibody products and services, and the prepared antibodies have strong specificity and good stability. Multiple antibody purification instruments and equipment can be used to provide antibody purification services from various sources such as rabbit, sheep, chicken, mouse monoclonal antibodies, as well as protein A/G affinity purification services and antibody separation services.

Based on various antibody preparation techniques such as hybridoma cell technology, Alpha Lifetech can provide customized services for monoclonal and polyclonal antibodies of different species. In addition, a range of services from high throughput sequencing, antibody preparation, antibody purification to antibody validation can also be provided. We will choose the purification method based on the specific purpose of the customer. Based on comprehensive platform systems such as antibody development platform and protein platform, Alpha Lifetech covers upstream and downstream services of antibody production, providing antibody purification, antibody high throughput sequencing, antibody validation and other technical services.

Hybridoma Technology

Hybridoma technology can be used to prepare monoclonal antibodies. By fusing spleen cells from immunized mice with myeloma cells from humans or mice, hybridoma cells are formed and secrete specific antibodies. The generated mouse derived monoclonal antibodies can be used in scientific research such as enzyme-linked immunosorbent assay, radioimmunoassay, immunohistochemistry, flow cytometry, protein purification, etc. Monoclonal antibody samples include ascites, hybridoma supernatant, etc. The sample obtained by injecting hybridoma cells into the abdominal cavity of animals and using the physiological environment of the animal's abdominal cavity to produce antibodies is called ascites sample. Hybridoma cells injected into the abdominal cavity of animals can proliferate and secrete monoclonal antibodies, which can accumulate in ascites. Due to the possibility of impurities (animal proteins, hormones, etc.) in these antibodies, purification is required. Based on the secretion function of B lymphocytes, monoclonal antibodies are often secreted into the culture medium when cultivating hybridoma cells. The liquid containing monoclonal antibodies in the culture medium is called hybridoma supernatant. The purity of monoclonal antibodies in the supernatant is relatively high, but further purification is required for large-scale preparation as they are obtained through in vitro culture.

Antibody purification can isolate high-purity antibodies from complexes. At present, antibody affinity purification and antibody separation and purification are commonly used antibody purification methods, mainly including protein A/G purification, ion exchange chromatography, gel filtration, precipitation and hydrophobic interaction chromatography. Among them, protein A/G affinity purification service is widely popular in various antibody purification applications. The process of Alpha Lifetech preparing monoclonal antibodies based on hybridoma technology is shown in Figure 1.

hybridoma technology-Alpha Lifetech
Fig.1 Process diagram for preparing monoclonal antibodies based on hybridoma technology. 

Antibody Drug Conjugate and Hybridoma Technology

Antibody drug conjugate (ADC) is a novel biological agent that chemically couples antibodies with cytotoxic drugs. Antibody drug conjugate combines the specific recognition ability of antibodies with the powerful killing effect of cytotoxic drugs, and precisely delivers cytotoxic drugs into tumor cells through antibodies, thereby achieving precise treatment. The antibody portion in ADC drugs is often prepared through hybridoma technology or other antibody production techniques, such as genetic engineering antibody technology. Hybridoma technology provides high-quality antibody raw materials for ADC drugs. ADC drugs and monoclonal antibodies prepared through hybridoma technology play important roles in fields such as cancer treatment. They can be used alone or in combination to enhance therapeutic efficacy.
Monoclonal antibodies prepared by hybridoma technology have high specificity, can accurately bind and recognize specific antigens, and can be produced on a large scale. Through hybridoma sequencing technology, antibody genes in hybridoma cells can be extracted and sequenced to obtain corresponding sequences, which is beneficial for subsequent antibody expression. Monoclonal antibodies prepared through hybridoma technology can be used to detect specific disease markers, such as tumor markers, viral antigens, etc., providing strong support for early diagnosis of diseases. In addition, it can also be used in experiments such as immunoblotting and immunohistochemistry to study protein expression, localization, structure, and function.

hybridoma technology Production Antibody Service Workflow

Steps Service Content Timeline
Step 1: Animal immunization
(1) Animal immunization 4 times, booster immunization 1 dose, a total of 5 doses immunized.
(2) Negative serum before immunization was collected, and ELISA was performed on the fourth dose to detect serum titer.
(3) If the serum antibody titer of the fourth dose meets the requirements, one additional dose of immunization will be administered 7 days before blood collection. If it does not meet the requirements, routine immunization will continue.
(4) Qualified potency, Spleen cells are isolated for cell fusion.
10-12 weeks
Step 2: Fusion and Screening
(1) Fusion: After immunization, mouse spleen cells were collected and fused with myeloma cells.
(2) Proliferation and screening: The obtained hybridoma cells were subjected to proliferation and ELISA screening to obtain highly efficient clones.
6 weeks
Step 3: Subclone
Subcloning 15-25 positive mother cells using limited dilution method, and finally determining 10-15 positive clones
3-4 weeks
Step 4: Ascites Production and Antibody Purification
(1) Ascites production: 10-15 hybridoma strains are used for ascites production in mice (if a large number of hybridoma strains are obtained, they will be sent in cellular form)
(2) Ascites identification: Ascites ELISA detection, potency (protein/virus>10 ^ 5, small molecule/peptide>10 ^ 4) for downstream purification
(3) Purification of Protein A/G: Purification of Ascites
3-4 weeks
Step 5: Delivery
(1) Provide 3-5 monoclonal antibody hybridoma strains. 
(2) Each hybridoma cell provides 1mg of purified antibody (ascites purification).
(3) Experimental Process Report
1 week

Advantage Of Hybridoma Technology

Alpha Lifetech has rich experience in the field of antibody development. Over the years, Alpha Lifetech has established a comprehensive antibody development platform.

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Multiple Species

Immunity can be provided to various animals, including but not limited to mice, rabbits, sheep, etc

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Multiple Target

Based on hybridoma technology, multiple target antibody discovery services are available: proteins, peptides, small molecules, viruses, membrane proteins, mRNA, etc. 

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Traceability Records

QC quality control standards (immune potency, PBMC quality control, library quality control, and screening validation quality control), English experimental reports, original experimental records

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Personalized Program

One on one personalized program customization  to meet the research project needs of various clients

Related SERVICE

Multiple Antibody Development Strategies

monoclonal antibody development-Alpha Lifetech

Monoclonal Antibody Development Service

We can provide high-quality, high-purity, and highly specific monoclonal antibody development services, including the production of mouse monoclonal antibodies and rabbit monoclonal antibodies

hybridoma cells-Alpha Lifetech

Hybridoma Technology Platform

Including immunization program, antibody preparation services, antibody purification, antibody high throughput sequencing, antibody validation, etc

b cell screening-Alpha Lifetech

Single B Cell Sorting Platform

Alpha Lifetech has advantages in screening time and obtaining high-quality antibodies. It can provide antigen design, synthesis, and modification, animal immunity, single B cell enrichment screening, single cell sequencing.

phage display-Alpha Lifetech

Phage Display Antibody Development Platform

Alpha Lifetech can provide phage display antibody development technical services from antibody preparation, antibody purification, antibody sequencing, etc.

Related Technology Resource

Introduction to Hybridoma TechnologyVIDEO

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