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    Surface Display

    Ribosome Display 

    Syd Biotechnologies specialises in the construction and screening of ribosome display libraries. These libraries, composed of diverse peptide sequences, cDNAs, and antibodies, are engineered with specific elements such as T7 RNA polymerase promoters, ribosome binding sites, actinomycin/puromycin interaction sites, and sequences to enhance RNA stability. Following in vitro transcription, translation is carried out using E. coli cell lysates, tRNA, buffer systems, and elevated Mg²⁺ concentrations at 4°C. Target–ligand–mRNA–ribosome quaternary complexes are then selectively captured and enriched through three to four iterative screening rounds. The RNA from these complexes is extracted, reverse transcribed, and cloned into a suitable vector. Finally, the resulting clones are sequenced and translated to determine the amino acid sequences of the selected ligands.  

    Service Highlights 

    • Libraries with 10^13-15 diversity  
    • Library from immunized animals  
    • Peptide library of different lengths  
    • cDNA libraries  
    • Specially designed libraries  

     mRNA Display Services  

    Syd Biotechnologies specialises in the development and screening of mRNA display libraries. These libraries, comprising a diverse array of peptide sequences, cDNAs, and antibodies, are engineered with essential features including T7 RNA polymerase promoter sites, ribosome binding sequences, actinomycin/puromycin binding regions, and RNA stability elements. 

    The workflow begins with in vitro transcription, during which mRNA is synthesised and covalently linked to puromycin. This linkage facilitates the stable association between the mRNA and the corresponding translated protein. Translation is performed in vitro using E. coli or eukaryotic cell lysates, in the presence of tRNA, buffers, and elevated concentrations of Mg²⁺. 

    Following translation, the resulting polypeptide–mRNA–puromycin complexes are purified and exposed to the target molecule. Through multiple rounds of enrichment, target–peptide–mRNA–ribosome quaternary complexes are selectively isolated. RNA from these complexes is then extracted, reverse transcribed, and cloned into a vector for sequencing. The amino acid sequences of the binding ligands are subsequently identified through translation. 

    Utilising this ribosome display platform, Syd Biotechnologies is capable of screening ultra-diverse libraries—reaching up to 10¹⁴ to 10¹⁵ variants—greatly enhancing the probability of discovering high-affinity binders for a wide variety of biological targets.  

    Service Highlights 

      We generate and screen highly diverse libraries with complexities exceeding 10¹⁴–10¹⁵ variants. 

      Antibody libraries derived from immunised animals are constructed and screened for target-specific binders. 

      Peptide libraries of varying lengths—including mini-proteins and small peptides—are developed and evaluated. 

      We offer cDNA library preparation tailored for mRNA display platforms. 

      Custom libraries incorporating non-standard amino acids and cyclised peptides are also available. 

      Selected clones are validated using techniques such as FACS, ELISA, and Biacore for binding specificity and affinity. 

     

    Phage Display Surface Display  

    Syd Biotechnologies offers premium custom phage display library construction and screening services to advance biological research and drug discovery initiatives. By introducing random mutations into the gene of interest, we generate a diverse array of peptides, small antibodies (such as scFv and Fab), or proteins, which are displayed as fusion proteins on the surface of filamentous phages (e.g., M13, fd, or f1 strains). 

    Using a binding affinity-based method known as panning, phages that present proteins with specific affinity for a target molecule are selectively enriched from a library comprising billions of unique variants. The identities of these target-binding proteins are then determined through phage amplification followed by DNA sequencing. 

     

    Our Approach  

    Syd Biotechnologies employs the yeast display system to monitor antibody maturation. Antibodies expressed on the yeast cell surface are exposed to antigens under progressively stringent conditions. Yeast cells displaying strong antigen-binding are then isolated using Fluorescence-Activated Cell Sorting (FACS). The selected antibodies are subsequently purified using affinity chromatography. 

     

    Service Highlights 

      Construction of phage display cDNA libraries tailored for specific research applications 

      Development of phage display peptide libraries with diversity ranging from 1×10⁸ to 1×10¹² variants 

      Library screening through multiple rounds (typically four) of panning using affinity chromatography and phage amplification 

      Screening of NEB’s Ph.D.™-7, Ph.D.™-12, and Ph.D.™-C7C peptide libraries 

      Screening and selection from custom phage display cDNA libraries 

      Bacterial expression and purification of scFv and Fab antibody fragments 

      Conversion of scFv and Fab fragments into humanised IgG antibodies, followed by expression in mammalian cell systems 

     

    Yeast Display 

     

    Syd Biotechnologies offers comprehensive yeast display library construction and screening services. Yeast display (also known as yeast surface display or YSD) is a powerful technique that enables the expression of proteins on the surface of yeast cells following proper folding and post-translational modifications within a eukaryotic system. Compared to phage display, yeast display offers significant advantages for the presentation of proteins and antibodies, as the eukaryotic environment of yeast allows for more accurate protein folding, modification, and translocation before surface display. 

    Libraries of human antibodies displayed on yeast are screened using flow cytometry, ensuring that the antibodies maintain conformational and functional properties closely resembling their native human forms. 

    Our scientists have over 20 years of experience and have developed a proprietary transformation protocol that consistently yields up to 2×10⁸ transformants per microgram of ligation mixture. This enables the generation of highly diverse and efficient libraries, making yeast display an excellent platform for high-affinity antibody discovery and affinity maturation.  

    Our Approach 

    Syd Biotechnologies utilises the yeast display system to monitor antibody maturation. Antibodies displayed on the surface of yeast cells are exposed to antigens under progressively stringent conditions. Yeast cells exhibiting strong antigen binding are isolated using Fluorescence-Activated Cell Sorting (FACS). The selected antibodies are subsequently purified through affinity chromatography.  

    Service Highlights  

    • Therapeutic antibody discovery  
    • Monoclonal antibody discovery  
    • Antibody Humanization  
    • Protein-protein interaction  
    • Antibody Affinity Maturation 

     

    Bacterial Display 

    Syd Biotechnologies offers the construction and screening of custom bacterial display libraries comprising small peptides. These libraries are valuable tools for ligand discovery, studying antibody–antigen binding affinities, and identifying novel molecular targets. 

    Polypeptides are displayed on the surface of bacterial cells and screened using flow cytometry or standard selection techniques such as biopanning. The libraries are generated using a proprietary method that combines a highly diverse collection of synthetically randomised peptide sequences. The system is specifically optimised to eliminate premature stop codons and aggregation-prone sequences, enhancing library quality and performance. 

    Similar to phage display, the peptides are expressed on the bacterial surface as fusion proteins anchored to the plasma membrane, with expression controlled through an inducible system. 

    We also accommodate customer-supplied vectors, provided the customer retains exclusive rights to the construct. 

     

    Our Approach 

     

    Syd Biotechnologies employs the yeast display system to monitor and guide antibody maturation. Antibodies displayed on the yeast cell surface are exposed to their target antigens under progressively stringent binding conditions. Yeast cells that exhibit high-affinity binding are selectively isolated using Fluorescence-Activated Cell Sorting (FACS). The selected antibodies are then purified using affinity chromatography for further analysis and validation. 

     

    Service Highlights: 

      Achieve high-level, uniform, and broad randomisation of peptides (8–30 amino acids) using strategies such as NNK codon schemes, split-mix-split synthesis, or tri-mer codon approaches. 

      Perform codon optimisation tailored to the host expression system to ensure efficient display of peptide sequences encoded by the inserted DNA. 

      Incorporate internal control tags (e.g., cMyc) and purification tags as per specific project requirements. 

      Purify oligonucleotides using polyacrylamide gel electrophoresis (both denaturing and native) to eliminate truncated inserts, enabling the construction of libraries with diversity ranging from 10⁹ to 10¹⁰ unique clones. 

     

    Global Protein Stability 

    Syd Biotechnologies offers global protein stability analysis using advanced yeast surface display technology. This platform enables the assessment and engineering of the stability of large protein libraries in a single, high-throughput experiment. 

    The process begins with in-silico design of engineered peptide sequences. These genes are synthesised, cloned into the pSYDYD 3.0 yeast display vector (or project-specific vector), and simultaneously expressed on the surface of yeast cells. 

    Displayed peptides are then challenged with various destabilising agents—including proteases like trypsin and chymotrypsin, shifts in temperature and pH, as well as denaturants such as urea and guanidinium hydrochloride. High-throughput screening enables the identification of protein variants with significantly enhanced stability. 

    This approach supports the development of more stable and functionally robust proteins for a range of applications, including therapeutic antibody development and industrial enzyme optimisation. 

     

    Service Highlights 

    • Optimized yeast display vector.  
    • High-throughput screening.  
    • Eukaryotic environment.  
    • Direct analysis on cell surface.  
    • Highly applicable in protein engineering.  
    • All services under one roof.  

     

    Experiment Steps: (Please regenerate the image)  

    Custom PhIP-Seq Service for Profiling of Autoantibodies 

    Syd Biotechnologies offers Custom Phage ImmunoPrecipitation Sequencing (PhIP-Seq) services for comprehensive profiling of autoantibodies present in patient serum, leveraging advanced phage display technology. 

    This platform utilises a phage display library constructed from predesigned oligopeptides ranging from 50 to 90 amino acids, cloned into a modified T7 Select vector developed in-house at Syd Biotechnologies. The library is then screened using antibody pools isolated from patients with suspected or confirmed autoimmune disorders. 

    Following incubation, the antibody–phage complexes are selectively captured using Protein A/G resin. A two-step PCR process is employed: the first round amplifies and purifies the phage-insert sequences; the second round adds unique index barcodes to differentiate the samples. Sequencing is performed using the Illumina MiSeq platform. 

    Post-sequencing, FastQ files are demultiplexed, aligned, and analysed to determine enrichment patterns of patient-specific autoantibodies. PhIP-Seq is a powerful tool for investigating environmental exposures, allergies, emerging autoimmune conditions, and patient responses to immunomodulatory therapies. 

     

    Highlights:
      Ready-to-Go modified T7 Select phage-display vector is available for immediate library construction.  

      One-Stop CRO for designing, library construction, immunoprecipitation, and NGS services.  

      Custom Vector modification project is accepted.  

      Fast turnaround time with approximately 2-3 weeks for library construction, 1 week for immunoprecipitation and 1-2 weeks for NGS.  

      Project customization and technical guidance are available throughout the project.  

      Gene synthesis, cloning, and library construction.  

      Screening of antibody pool by Immunoprecipitation using protein A/G resin.  

      NGS and analysis.  

      Validation Experiment. 

     

    Services Offered:
      Gene synthesis, cloning, and library construction.  

      Screening of antibody pool by Immunoprecipitation using protein A/G resin.  

      NGS and analysis.  

      Validation Experiment. 

     

     

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