By integrating the most comprehensive computational biology tools and databases, Creative Biolabs has discovered a series of solutions for designing scFv antibodies. We tailor the optimization of antibody affinity and stability to suit your specific research needs. Our high-throughput screening platform enables us to rapidly identify the scFv fragments with the best expression levels. Through a variety of biological assays, we assess the activity and binding characteristics of the antibodies, thereby accelerating your research and development process.

Introduction

Creative Biolabs is a leading biotechnology company focused on preclinical molecular discovery of antibody drugs. Our research and development team possesses extensive experience in antibody fragment development and utilizes a proprietary scFv recombinant antibody technology platform. Our platform works seamlessly with a functional protein expression system, enabling us to provide high-activity functional proteins. Our scFv recombinant monoclonal antibody development platform is designed for antibody development across various species, significantly reducing the time required compared to traditional hybridoma techniques. scFv antibodies produced through our platform exhibit excellent drug-like properties, and a high number of positive clones, and allow for direct access to antibody sequences, facilitating large-scale production of therapeutic target antibodies for downstream preclinical studies.

Fig.1 The Production of GD2-specific Antibody Fragments.^1,3

Services

ScFv Antibody Fragment Expression Solutions

At Creative Biolabs, the production of scFv recombinant antibodies can be approached through three main methods: phage display screening, sequencing of existing hybridoma cells, and direct B cell cloning. Our system not only produces high-quality scFv antibodies but also provides the genetic sequences of these antibodies, facilitating subsequent antibody production and modification.

1. Phage Display Screening Service

Phage display technology is a highly effective method widely used in antibody development, utilizing phages as a vehicle to present exogenous antigens. We begin by constructing a diverse scFv library, which is then cloned into the phage genome. Specific scFvs that bind to the target antigen are selected through binding assays, followed by elution and amplification. The successfully isolated scFv clones undergo further affinity testing and optimization to yield high-performance recombinant antibodies. Our approach ensures high throughput, excellent specificity, and relatively low costs, making it ideal for applications requiring rapid screening of specific antibodies.

2. Hybridoma Cell Sequencing Service

Our laboratory offers a quick solution to obtain specific antibody gene information by performing single-cell sequencing on existing hybridoma cells. The process starts with isolating hybridoma cells from immunized mice, followed by whole-genome sequencing to identify B cells capable of producing specific antibodies. The sequences obtained are then utilized to reconstruct scFvs, which are expressed in an appropriate expression system. This method allows for the rapid generation of recombinant antibodies targeting known antigens, significantly shortening the development timeline, especially when related antibodies are already available.

3. Direct B Cell Cloning Service

Recently, the process of isolating and cloning B cells directly from immunized animals has emerged as an effective approach for developing scFv antibodies. The initial step in this service involves extracting spleen cells from immunized animals, such as mice or rabbits, and then using flow cytometry to select B cells that produce specific antibodies. Following this, single-cell PCR technology is employed to amplify both the heavy and light chains of the antibodies from these B cells. Subsequently, gene recombination techniques are utilized to combine these two chains, resulting in the formation of scFv. This method harnesses the native immune response generated within the animal to produce antibodies, thereby enhancing affinity and specificity, making it particularly suitable for projects that require highly specific and high-affinity antibodies.

ScFv Antibody Fragment Purification Solutions

Creative Biolabs offers efficient scFv purification solutions to meet diverse customer requirements. Our scFv purification solutions use advanced separation technologies and optimized processes to significantly enhance the efficiency and yield of antibody fragment purification. Our solutions incorporate a combination of specific affinity chromatography, ion exchange chromatography, and gel filtration techniques, ensuring high purity and activity of the target antibody fragments.

1. Protein A Affinity Chromatography

Protein A affinity chromatography is a well-established method for antibody purification, extensively used for the preliminary separation and enrichment of scFv fragments. This technique exploits the high affinity between Protein A and the antibody Fc region, allowing for quick and effective extraction of target antibodies from complex samples.

- High Purity: Effectively removes impurities, enhancing the overall purity of the antibodies.

- User-Friendly Process: The straightforward procedure facilitates large-scale production.

- High Throughput: Ideal for handling large sample volumes, significantly saving time.

2. Ion Exchange Chromatography (IEX)

Ion exchange chromatography separates molecules based on surface charge and is suitable for fine purification based on isoelectric point (pI). For scFv, adjustments can be made at different pH levels to optimize separation efficacy.

- High Resolution: Capable of separating closely related isoforms.

- Strong Flexibility: pH adjustments allow for effective handling of various scFv variants.

- Broad Adaptability: Suitable for a wide range of protein separation and purification tasks.

3. Gel Filtration Chromatography (GFC)

Gel filtration chromatography purifies proteins primarily based on molecular size. This method is effective for eliminating small molecule impurities or aggregates, ensuring the isolation of target scFv monomers.

- Gentle Conditions: Maintains the structure and functionality of scFv, making it ideal for highly active antibody fragments.

- High Throughput: Facilitates rapid separation of large sample sizes, enhancing yield and efficiency.

4. Affinity Chromatography

In addition to protein A, we provide a wide variety of customizable affinity chromatography techniques to suit specific binding ligands.

- High Specificity: Effectively captures target scFvs with minimal non-specific binding.

- Flexibility: Customized affinity matrices based on client requirements to achieve optimal purification strategies.

Fig.2 The Binding Analysis of scFv Antibodies.^2,3

Case Study

A. Project Background

Creative Biolabs is dedicated to developing therapeutic antibodies targeted at specific tumor markers. During the early stages of research and development, the client encountered challenges related to high costs and low yield of their antibodies. There was an urgent need to optimize the processes for antibody expression and purification. In response, our experimental team initiated a project aimed at enhancing the expression efficiency of ScFv antibody fragments and streamlining subsequent purification steps.

B. Project Implementation

Our team formulated a systematic experimental approach. We began by employing genetic engineering techniques to construct an efficient expression vector for the ScFv antibodies. Subsequently, we adopted optimized expression conditions, such as fine-tuning temperature and inducer concentration, to ensure the ScFv fragments could be expressed effectively in E. coli.

For purification, we introduced a novel affinity chromatography technique, complemented by our customized purification reagent kit, which significantly improved the separation and purification of ScFv antibodies. Throughout the project, we conducted multiple small-scale experiments, continuously adjusting and refining our methods until we identified the optimal protocol.

C. Project Outcomes

After several months of dedicated work, our project team achieved remarkable results:

• High Expression Levels: We successfully reached an expression level of 30 mg/L for the scFv antibody fragments in E. coli, greatly exceeding the client's initial expression levels.
• High Purity: The optimized purification process yielded scFv antibody fragments with a purity of over 95%, ensuring safety and biological activity for subsequent applications.
• Retention of Activity: Biological activity assays demonstrated that the purified scFv antibody fragments exhibited excellent affinity for their targets, with IC₅₀ values significantly lower than those observed before optimization.
• Reduced Timeline: Following the implementation of the project, the client's antibody development timeline was shortened by 30%, significantly accelerating their market entry.

Advantages

In recent years, recombinant antibodies have become increasingly prevalent in the field of biomedicine, serving as powerful tools for treating a variety of diseases. With ongoing advancements in technology, ScFv-Fc has emerged as a novel class of biotherapeutics, drawing significant attention in research and application. Creative Biolabs, a top preclinical CRO, is committed to providing efficient and precise ScFv Antibody Fragment Expression and Purification Services, helping our clients navigate the antibody drug development process with greater stability and success.

• Enhanced Affinity and Specificity: ScFv-Fc fusion antibodies maintain the high affinity of ScFv while benefiting from the stability and functionality provided by the Fc region. This makes ScFv-Fc particularly effective in targeted therapies and diagnostics, offering superior selectivity.
• Improved Biological Effects: The inclusion of the Fc region allows ScFv-Fc antibodies to more effectively trigger ADCC and CDC, thereby enhancing their therapeutic efficacy.
• Versatile Design and Application: The ScFv-Fc structure can be flexibly engineered to meet various research needs and application scenarios, making it widely applicable for treating diverse diseases such as cancer and immune disorders.

Creative Biolabs boasts a team of top-notch scientists capable of offering customized scFv antibody design services tailored to client needs. For the purification of scFv antibodies, we utilize advanced chromatography techniques, including affinity chromatography and ion exchange chromatography, which allow us to efficiently eliminate impurities and ensure high purity and functionality of the antibody fragments. In addition to providing expression and purification services for antibody fragments, we also offer a comprehensive range of functional assessment tests to assist clients in validating the properties and activity of the antibodies. If you would like to learn more about our services or inquire about project details, please feel free to contact us. We are here to provide you with support and assistance.

For research use only. Not intended for any clinical use.