Engineered Protein Scaffold Discovery Service

Background Workflow Features Various Scaffold Introduction

Scaffold proteins are characterized by the presence of specific protein-binding domains. To date, hundreds of human proteins have been defined as scaffold proteins. Based on the advanced trimer codon technology, Creative Biolabs can introduce mutations in specific domains of scaffold proteins to offer custom scaffold library construction and screening services, which greatly expands the scope of your research applications.

Introduction of Scaffold Protein

Scaffold proteins, as molecular hubs for protein docking, play an important role in various signaling pathways. In this process, scaffold proteins can correctly regulate intracellular signal transduction. The Fig. 1 A concept map of sequence design step of scaffold library construction. (Allen, et al., 2010) engineered scaffold proteins have properties similar to or even better than antibodies and are used in various biomedical and biotechnological applications. The rich protein skeletons in nature provide a valuable source for the development of engineered scaffold proteins. Creative Biolabs can offer custom scaffold protein library construction and screening solutions to allow you to work with the scaffold of interest.

Workflow

Fig. 2 A workflow of Creative Biolabs’ scaffold discovery process, including sequence design, library construction and library screening. (Creative Biolabs Original)

Scaffold Sequence Design

Fig. 3 Plasmid design for scaffold library construction. (Creative Biolabs Original)

Based on the selected parental scaffold sequence, we need to determine the domain to introduce mutations. Subsequently, the trimer codon technology was used to introduce mutations at these positions. In general, the mutant amino groups were the remaining 19 amino acids except Cys, distributed in equal proportions.

Scaffold Library Construction

Phage Display Library Construction

Based on the specific project objective and target information, we can provide the most suitable phage display system for scaffold protein library construction, including M13, T4, and T7.

Yeast Display Library Construction

Yeast display libraries are mainly used to screen proteins with specific binding properties. Yeast display provides a possibility when other display methods are not effective.

Scaffold Library Screening

Solid-Phase Screening

Solution-Sorting Screening

Cell-Based Screening

In Vivo Screening

Ex Vivo Screening

Advantages

Small size allows to penetrate cell membranes with high permeability.
Higher thermodynamic stability.
Lower aggregation.
Binds to multiple targets with high affinity and specificity.
Rigid protein framework for versatile engineering concepts.

How Scaffold Discovery Service Can Assist Your Project?

Scaffold discovery is the process of finding the most suitable scaffold protein binder for the client's target. First, we need to discuss with the client and determine the scaffold type to be used. Then we can build a scaffold mutation library with high precision and an expected size of over 10¹⁰. Through efficient library screening, we can finally obtain the most imaginative scaffold protein binders.

Various Scaffold Introduction

A B C E F G H I K L M N P S T W Z Others

A APP (Avian Pancreatic Polypeptide) APPI (Alzheimer's Amyloid β-Protein Precursor Inhibitor) Aptide Alphabodies

B BPTI (Bovine Pancreatic Trypsin Inhibitor)

C CBD (Cellulose-Binding Domain) CBM4-2 (The Second Family 4 Carbohydrate Binding Module) Charybdotoxin (CTX) CTLD (C-Type Lectin-like Domain) Cytochrome b₅₆₂

E Ecotin EETI-II (The Ecballium elaterium Trypsin Inhibitor II)

F FN3 (Fibronectin Type III Domain) Fynomers

G GCN4 GFP (Green Fluorescent Protein)

H Human Serum Transferrin (hTF) HEAT

I Im9 (The Immunity Protein 9) Insect Defensin A Intrabody

K Knottins

L LACI-D1 (Human Lipoprotein-Associated Coagulation Inhibitor Domain 1)LDTI (The Leech-Derived Tryptase Inhibitor)

M Min-23 Monobody MTI-2 (The Mustard Trypsin Inhibitor 2)

N Nanoantibody (nAb)NanoScaffold Neocarzinostatin (NCS)

P PBP (The Periplasmic Binding Protein) PDZ Domain PHD Finger (The Plant Homeodomain Finger) Protein A Scaffold PSTI (Pancreatic Secretory Trypsin Inhibitor) pVIII

S SFTI (Sunflower Trypsin Inhibitor) SH2 (Src Homology 2) SH3 (Src Homology 3) SPINK1 (Serine Protease Inhibitor Kazal-type 1) Staphylococcal Nuclease

T TATI (Tumor-Associated Trypsin Inhibitor) TEM-1 β-lactamase Tendamistat TFPI-I (Tissue Factor Pathway Inhibitor-I) Thioredoxin (TrxA) TPR (Tetratricopeptide Repeat) Trans-body

W WW Domain

Z Zinc Finger

Others α₂p8 αRep

Creative Biolabs is professional in applying advanced library construction and screening platforms for scaffold discovery. If you are interested in our service, please do not hesitate to contact us for more information.

Reference

Allen B D, et al.; Experimental library screening demonstrates the successful application of computational protein design to large structural ensembles. Proceedings of the National Academy of Sciences, 2010, 107(46): 19838-19843.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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