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Small molecule Immunization

Category : Immunization Service • Last Updated : 2026-02-08 12:23:19 • Hits : 337

Small-molecule immunization is the mainstream technique for generating small-molecule-specific antibodies. Small molecules with a molecular weight below 1000 Da act solely as haptens and cannot trigger an immune response independently. They must be covalently conjugated to macromolecular carrier proteins such as KLH and BSA via carbon-chain linkers to form complete antigens. The T-cell epitopes carried by the carrier then activate the host’s humoral immunity.

Small-Molecule Immunization Procedure：

No.

Time

Immunization Procedure

1

Day 0

Collect 10 mL of whole blood as reserved negative serum.

2

Day 1

1st: 0.5 mg small-molecule immunogen + Complete Freund's Adjuvant (CFA); Subcutaneous injection at 10 sites.

3

Day 14

2nd: 0.25 mg small-moleculeimmunogen + Incomplete Freund's Adjuvant (IFA); Subcutaneous injection at 5 sites.

4

Day 21

Collect 1 mL of whole blood, separate serum, and perform ELISA titer testing.

5

Day 28

3rd: 0.25 mg small-moleculeimmunogen + Incomplete Freund's Adjuvant (IFA); Subcutaneous injection at 5 sites.

6

Day 35

Collect 1 mL of whole blood, separate serum, and perform ELISA titer testing.

7

Day 42

4th: 0.25 mg small-molecule immunogen + Incomplete Freund's Adjuvant (IFA); Subcutaneous injection at 5 sites.

8

Day 49

Collect 10 mL of whole blood, separate serum, and perform ELISA titer testing. Based on titer results:

  • Collect 50 mL of whole blood, separate serum, perform protein ELISA titer testing. 
  • From the remaining whole blood, isolate PBMCs, lyse 2.0E+07 cells with 1 mL TRIzol, and aliquot at 1 mL per tube.

Result Analysis：


Deliverables：

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