Unlock instant, AI-driven research and patent intelligence for your innovation.

Additive used for increasing clone number of hybridoma cells and preparation method of additive

A technology of hybridoma cells and additives, which is applied in the direction of cell culture active agents, biochemical equipment and methods, fusion cells, etc., can solve the problems of unfavorable animal welfare and expensive hybridoma additives, and achieve low cost and low economic cost. Cost and labor cost, the effect of animal protection

Active Publication Date: 2016-04-06
同昕生物技术(北京)有限公司
View PDF1 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when preparing feeder cells, 4-5 mice must be killed for each fusion, which is not conducive to animal welfare, and the price of commercial hybridoma additives is relatively expensive

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Additive used for increasing clone number of hybridoma cells and preparation method of additive
  • Additive used for increasing clone number of hybridoma cells and preparation method of additive
  • Additive used for increasing clone number of hybridoma cells and preparation method of additive

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1, be used to improve the preparation of the additive of hybridoma clone quantity

[0026] 1. Design and synthesis of primers

[0027] According to the known Musmusculus interleukin6 (I16) sequence (GeNBank No. GeneID: 16193), the sequence was optimized with JavaCodonAdaptationTool software to obtain the sequence IL-6* (as shown in SEQ ID NO.: 1). The sequence was synthesized by Shanghai Sangong and cloned into the pGH plasmid to obtain the pGH-IL-6* plasmid. Its PCR primers are:

[0028] Upstream primer IL-F: 5'-AAGGTACCatgaaattcctgtctgctcg-3',

[0029] Downstream primer IL-R: 5'-TATCTAGAttaggtctgacgggtag-3'

[0030] The upstream primer and downstream primer have KpnI and XbaI restriction sites respectively. The primer was synthesized by Shanghai Sangong.

[0031] 2. Construction of eukaryotic expression vector pcDNA5 / FRT-IL6

[0032] Using the pGH-IL-6* plasmid as a template, the IL-6* sequence was amplified by PCR, and the sequence was double-digeste...

Embodiment 2

[0040] Embodiment 2, for improving the effect measurement of the additive of hybridoma clone quantity

[0041] 1. Preparation of hybridoma cells

[0042] (1) Animal immunity

[0043] 50 μg of BSA protein was diluted with 100 μl of 20 mM phosphate buffer (pH 7.4) containing 0.15 M NaCl, and mixed with an equal amount of Freund's complete adjuvant to make an emulsion. Inject subcutaneously at multiple points in 8-12 week old BALA\C mice. After 3 weeks, take the same amount of antigen and incomplete adjuvant for booster immunization once; after another 4 weeks, take the same amount of antigen without adjuvant for intravenous booster immunization.

[0044] (2) Preparation of immune spleen cell suspension

[0045] On day 3 after the last immunization, the immunized mice were sacrificed. The spleen was removed in an ultra-clean workbench and placed on a 100-mesh stainless steel screen, gently ground with a syringe core, washed with serum-free medium, and the cells were collected...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides an additive used for increasing the clone number of hybridoma cells and a preparation method of the additive, and relates to an immunological research technology. The preparation method includes the following steps that 1, culture solution supernatant for eukaryotic cells capable of expressing IL-6 protein is acquired, wherein the cell culture solution supernatant contains expressed IL-6 protein; 2, the cell culture solution supernatant is subjected to ultrafiltration and filtration sterilization in sequence, and a cell culture solution supernatant concentrated solution is obtained; 3, sodium selenite is added into the cell culture solution supernatant concentrated solution, and the additive is obtained. As proved by experimental data, the clone number of the hybridoma cells can be increased by nearly 5 times after the additive is adopted. The invention further provides a hybridoma cell culture medium containing the additive.

Description

technical field [0001] The invention relates to an additive for increasing the number of clones of hybridoma cells and a preparation method thereof, belonging to the fields of biology and medicine. Background technique [0002] Immunology is an integral part of today's medical and scientific research. Antibodies have been used in many ways since Edelman and Porter first isolated the immunoglobulin molecule 40 years ago. Antibodies are used in proteomics and diagnostics to detect tumor and bacterial antigens; they can also be synthesized in goat milk and plants as vaccines and anti-tumor factors; they can also be used as drug delivery tools to treat viruses, bacterial infections and tumors. [0003] Hybridoma technology and the production of monoclonal antibodies have revolutionized research, providing countless and unique monoclonal antibody reagents for the identification, isolation, clearance, activation or detection of specific antigens. Generation of antibody-secreting...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N5/20
CPCC12N5/163C12N2501/2306
Inventor 张建珍李全吴凡焦守恕
Owner 同昕生物技术(北京)有限公司