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Antibody Affinity Maturation Service

Antibody Affinity Maturation ServiceIntroduction

KMD Bioscience has been dedicated to antibody research for many years. Antibody affinity is a core parameter that determines the binding activity, potency, and clinical application potential of antibody drugs. Humanized antibodies generated via CDR grafting often suffer from varying degrees of affinity reduction due to conformational changes in antigen binding caused by framework region sequence substitutions. Therefore, directed improvement of antibody affinity through in vitro affinity maturation technologies represents a critical step in the antibody optimization phase.

KMD Bioscience possesses a well-established phage display platform and affinity characterization system, offering one-stop affinity maturation services for antibodies derived from multiple species. We are capable of constructing mutant libraries with a diversity exceeding 1010 independent clones. Through rigorous multi-round screening and identification, most projects achieve an affinity improvement of 1–3 orders of magnitude, reaching nanomolar or even picomolar levels. In parallel, we provide comprehensive support including antibody sequence analysis, soluble expression, affinity and specificity validation, and customized optimization strategies tailored to individual client needs, efficiently supporting early-stage development of antibody therapeutics.


Phage Display Antibody Library Technology - Antibody Affinity Maturation Service

In vivo affinity maturation is a natural evolutionary process of humoral immunity. During the germinal center response, antigen‑activated B cells undergo somatic hypermutation in the antibody variable regions, particularly within the complementarity‑determining regions (CDRs). Through continuous antigen‑driven selective pressure, B cells expressing high‑affinity antibodies are preferentially expanded, ultimately leading to a progressive increase in antibody affinity.

KMD Bioscience leverages phage‑displayed antibody library technology in combination with multiple mutagenesis strategies, including error‑prone PCR, site‑directed mutagenesis, and CDR‑directed saturation mutagenesis, to precisely recapitulate the "mutation‑selection‑enrichment" evolutionary process in vitro, thereby enabling directed acquisition of high‑affinity antibody variants.


Antibody Affinity Maturation Service Workflow

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Common Antibody Affinity Maturation Techniques in Vitro

During in vitro Antibody Affinity Maturation, selecting the mutation region and the method for introducing mutations are critical issues. Current mutation strategies are primarily categorized into three classes: Random Mutagenesis, CDR Mutagenesis and Site-Directed Mutagenesis, and Error-prone PCR.

Type

Characteristics

Error-Prone PCR

* This is a commonly used antibody mutagenesis technique that can randomly introduce mutations throughout the entire antibody gene or within specific regions. During polymerase amplification of the target gene, mutations are randomly introduced at a certain frequency by employing low-fidelity polymerases or adjusting reaction conditions. Multiple rounds of PCR are performed to iteratively introduce random mutations, accumulating the mutational effects, ultimately yielding random mutants of the target protein.

Chain Shuffling

* Chain shuffling involves retaining either the heavy chain or light chain of a specific antibody, while pairing it with a randomized complementary chain. Higher-activity mutants are then selected through screening of the phage antiboyd library.

Site-directed Mutagenesis

* Somatic hypermutation occurs predominantly in the CDR regions, which directly contact the antigen. During in vitro antibody affinity maturation, the CDR regions are frequently chosen for site-directed mutagenesis. When performing site-directed mutagenesis on CDRs, multiple CDRs can be mutated in parallel or optimized sequentially.

DNA Shuffling

* DNA Shuffling Technology involves cleaving homologous antibody genes into fragments not exceeding 50 bp using DNase I. These fragments are then randomly reassembled and amplified by PCR into full-length antibody genes.

Delivery Result

Delivery Content

Affinity-Optimized Antibody Sequences

High-Affinity Purified Antibodies

Complete experimental reports and data: including results such as SDS-PAGE and affinity analysis.


Service Advantage

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