Modern Procedures of Antibody Sequencing
Sep 15th 2022
Investigating the genetic makeup of proteins and antibodies is crucial for applications such as bioengineering, medical development, and other scientific studies. Whether you are involved in small-scale or large-scale genetic studies, you need the service of independent third-party laboratories to help with different needs. Shifting such burdens enables you to focus on the main study and boost the quality of the results.
Genetics research laboratories focus on helping individual and organizational researchers get all the needed genetic materials and items. They offer services such as antibody sequencing, gene synthesis, and production. These services are critical to helping you get accurate results, the suitable samples you need, and modify different genetic outcomes. Here is a guide to some of the services offered.
Sequencing services
The labs offer a wide range of sequencing services based on your needs, technology, and anticipated results.
Monoclonal sequencing
Most antibody samples are described based on the 'clonality,' which refers to the number of sequences or lineages available in the sample. In Monoclonal sequencing, the samples share the same genetic sequencing. In this approach, monoclonal antibodies undergo proteomic sequencing. There are different types of monoclonal sequencing. Monoclonal sequencing differs from polyclonal sequencing. In polyclonal sequencing, the antibody samples have diverse antibody sequences and contain a wide range of antibodies from the serum. Common monoclonal sequencing services and approaches include:
Edman sequencing
The approach varies from others due to the technology used. This approach is ideal for determining protein sequencing and reading one amino acid at a time, beginning with the N-terminus. This method is ideal for custom protein synthesis since it requires low material requirements. However, it is not ideal for full-length antibody sequencing since the amino acids degrade as you perform another sequencing.
The method enables the labs to identify the peptide sequence of an antibody. It involves repeatedly removing a single amino acid, identifying it then repeating the process to analyze all the amino acids. This method can sometimes be slow due to its repetitive nature.
Mass spectrometry-based sequencing
This method is popular due to the full-length antibody sequencing. The genes undergo peptide synthesis through the mass spectrometer. The peptides used will result in fragment spectrum; hence the laboratories must independently interpret every fragment spectrum to boost the antibody sequencing methods.
Peptide sequencing services
Peptide sequencing services mainly rely on the De Novo approach and technology. The procedure involves the use of an unknown sequence. The process requires the digestion of few enzymes and fewer mass spectrometry than in the De Novo technologies hence lowering the price for the whole process.
De Novo technologies are ideal for the sequencing process due to the ability to sequence the amino acid sequence of a custom antibody. The process begins by fragmentation of the antibody before subjecting it to mass spectrometry which is necessary for identifying the spectrum of mass to charge ratios in each fragment. The sequence is significant to determine and reconstruct the fragmented sequence into one total sequence using technologies such as bioinformatics software to identify any overlapping sequence.
De Novo Antibody Protein Sequencing Services involves different approaches using different sequencing technologies. These approaches include
Shotgun protein sequencing
This custom protein synthesis method merges overlapping sequences to read and identify the full-length chromosomes. This method involves sequencing all the fragments except those produced by a mass spectrometer. This method is not ideal if you require high levels of accuracy, especially when you need a single-spectrum analysis. This method is ideal for avoiding the genetic synthesis of a single-spectrum fragrance.
Templates proteogenomics
The peptide synthesis method is less suitable for complete antibody sequencing since some are unique and generally unknown before the primary sequencing method. The large portions of the antibodies are unmutated from the sequencing and can confidently be determined using peptide synthesis.
Small-scale sequencing services
Sequencing services are also available for small-scale services for independent studies, experimentation, and other scientific needs. One of the small-scale approaches is the Sanger sequencing. This method divides the DNA into four vials with DNA polymerase and four deoxynucleotides. The next stage is transcribing the DNA in each vial by adding modified di-deoxynucleotides; triphosphates (ddNTPs) to terminate DNA elongation. The generated fragments undergo different electrophoresis to determine the DNA sequencing related to the brand.
Large-scale sequencing services
Large-scale sequencing services are diverse and mainly target big corporations such as drug manufacturers, research institutions such as CDC, and those involved in genetics. Genetics labs heavily rely on AI and other technologies to boost the quality and speed of services. The technologies are ideal for identifying nucleic acid DNA sequencing. These methods use different technologies to produce faster results and conduct multiple sequencing in parallel.
The process also undergoes different stages to produce the desired results. The process involves four steps mainly:
- Library preparation: this stage involves DNA fragmentation and adding adapters to each end.
- Cluster amplification: The fragments are hybridized and then multiplied.
- Sequencing: In this stage, the labs add reagents to the custom antibody as the sequencing begins. The antibodies release different emissions and different specific wavelengths.
Why outsource peptide sequencing services?
Antibody sequencing services include different approaches and the utilization of different technologies, which you may not have hence the need to outsource. Third parties have technologies suitable for processes such as sequencing transcripts and genetic materials from a custom antibody from different serums or B-Cell.
Confidentiality: If you want to analyze any custom antibody, you can rely on the labs to keep the results secret and only release them to you. Most labs store the sequences on stand-alone private servers; hence the knowledge is unavailable to the public.
Flexibility: You can easily access independent antibody production services for different studies and experiments. The antibodies used are from different animals, such as mice, rats, and rabbits, while complying with various regulations in antibody production. You can also get other variations to boost studies' and analysis accuracy or dependability.
Accuracy: Leading genetics laboratories have the relevant technologies and materials to help with genetics synthesis. The technologies are ideal for eliminating repetition or specific processes that may increase errors.
Speed: High-tech labs usually need about two to four weeks to complete the services. They utilize many efficient technologies and procedures for processes such as mRNA purification, cDNA generation, and targeted PCR to boost the result accuracy while reducing the time needed to complete the whole analysis.
Antibody production services
There are different Custom antibody services for other clients based on the analysis they wish to carry out. The primary services include:
- Hybridoma-based monoclonal antibody production: The process involves producing monoclonal antibodies against non-modified, acetylation-specific peptides and other small compounds needed for experimentations such as ELISA, IHC, etc.
- Rabbit monoclonal antibody production service: The process involves the production of rabbit custom monoclonal antibodies against non-modified, phosphor-specific, acetylation-specific peptides, methylation-specific, and proteins suitable for IP, IF, IHC, and ELISA activities. The antibodies are ideal for different and wide ranges of targets such as small molecules, Ilama, antibody idiotype, etc.
- Human and monkey monoclonal antibody production service: For such services, you need top laboratories with a monkey and human monoclonal antibodies developed through single B cell-based antibody generation technologies. You can also consider laboratories offering services such as Camelid monoclonal antibody production
Peptide synthesis services
Synthesized peptides are suitable for applications such as drug testing, designing enzymes, and the creation of antibodies. Rather than generating your antibodies, you can outsource the services due to the advanced technologies and equipment to boost quality results. This will enable you to focus on your core duty. The peptide synthesis methods involve two approaches:
- Solid-phase: This method is suitable for separating the peptide from the reaction mixture and protecting the C-terminal by attaching the solid resin.
- Liquid-phase: Liquid-phase involves synthesis in a solution to ensure the final products have the highest purity and boost convergent synthesis. The ideal method should meet your desired goals and objectives.
Custom peptide synthesis methods involve many peptide modifications, including amidation, acetylation, methylation, etc. Most of these activities are technology driven, hence high quality and faster turn-around services. You also get different purity levels based on the experiment you are conducting. The custom peptide synthesis includes various services:
- Standards peptides: These are ideal for basic research or biomarker discovery. There are over 400 modifications available with a turn-around period of 3 to 4 weeks, depending on quantity.
- Alexa Fluor Dye-Labeled Peptide Synthesis Service: This is suitable for fluorescence imaging. The synthesis method involves the use of high-performing dyes to boost the quality.
- PEPotec Immuno Peptide Libraries are suitable for vaccine development and epitope mapping. They have low toxicity and high throughput.
- PEPotec SRM Peptide Libraries: These peptides are suitable for discovery due to the high throughput.
- Heavy Peptide AQUA Peptide Synthesis Service: They are ideal for large-scale operations such as quantitative mass spectrometry. The results are accurate due to the high precision and quality.
Gene synthesis services and protein expression services
Gene synthesis is becoming an integral part of modern biological studies and research. The DNA synthesis process involves producing gene-length DNA strands for different purposes. The process is complicated and delicate; hence you need to hire one of the most reliable Contract Research Organizations (CROs) to help you with DNA sequencing, protein expression, genotyping, and purification.
The turn-around duration for custom Gene Synthesis varies based on quality and the technology used. The duration should be 5 to 25 days. The process involves subjecting all the genes through a standard process that involves bioinformatics, oligo synthesis, contraction, quality control, and delivery. This standard procedure will take about five days; however, it can also increase due to the add-on services. Gene synthesis also includes the following add-services:
- Cloning: The process involves cloning any gene sequence into the required vector.
- Mutagenesis: The services include base deletion, insertion, site-directed mutagenesis, and arbitrary editing.
- Plasmid preparation: This step is necessary to prepare high-quality plasmids in varying scales from milligrams to grams.
- Protein expression and purification: This process can help you boost protein purity using world-class purification technologies if you need high-potent proteins.
Gene synthesis techniques
You can get different gene synthesis techniques based on other technologies and oligonucleotides. The first process involves the production of oligonucleotides to generate the required enzymes' building blocks. Labs can deploy different techniques such as
- Codon optimization: The platform mainly deals with next-generation gene syntheses, synthetic DNA libraries, vector construction, small genome syntheses and assembly, and pathway synthesis and assembly.
- PCR-Based Reactions: The Polymerase Chain Reaction method is one of the most common gene synthesis methods. It involved creating a DNA strand similar to the provided strand. The method is viable for commercial processes such as sequential chain reaction (SCR).
- Slonomics technology: The method relies on the universal DNA strand as a building block. This method is fully automated using industrial robots to minimize potential errors. This makes it superior to other methods due to the technology involved.
- The phosphoramidite method involves adding a 5' hydroxyl terminus to the nucleotides to form the sequential oligonucleotide chain. It involves capping, oxidation, DE protection, and coupling to boost the gene quality. Each process has an independent and integral role in boosting the final results.
- Chip-based technology: This process involves building a new gene based on a computer chip. It relies on different electrochemical and heat-controlled technologies.
ELISA test Kits
Elisa test kits are essential elements for companies such as those involved in logistics and transportation. There are different and high-quality ELISA test kits for different purposes. Before purchasing the kits, you need to verify the sources and ensure you get them from the best Elisa kit manufacturers. The manufacturer should have generic and modified kits based on your needs. A good manufacturer should deliver the kits in time for the tests. Based on the required designs and quantity, they should deliver between 3 and 11 business days.
You also need to consider the quality of the kits. A good Elisa kit should have the following features:
High sensitivity and low detection range: they can test for drugs and other elements in low dosage or range. The detection ranges are (1-100pg/mL, 3-200pg/Ml) making them highly accurate.
Sample types: The kits should be suitable for various samples such as serum, plasma, cell culture supernatants, and cell lysates.
Specifications: The period of use varies depending on the kit you buy. The applications are also varied, such as shortened experimental times, pre-collated detection reagents
Customer support: The manufacturer should offer the best products you need for your upcoming analysis. They should have the best return policies and ship the products within the shortest duration.
The Final Verdict
Many genetics laboratories are offering various services for different scientific and research purposes. You must ensure they provide quality solutions using state-of-the-art technology and products to boost the results and turn-around period. Since the modern procedures of antibody sequencing comprise various technologies and methods, you must be vigilant to pick the right one.
Links
- https://inpeaks.com/2020/04/24/methods-and-applications-of-gene-synthesis/
- https://www.antibodies-online.com/resources/17/5034/peptide-synthesis-methods-and-reagents/
- https://www.sinobiological.com/resource/antibody-technical/antibody-production
- https://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0218717
- https://www.biomatik.com/
- https://pubmed.ncbi.nlm.nih.gov/31233538/
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2891972/
- https://www.mcponline.org/article/S1535-9476(20)31840-5/fulltext
- https://www.researchgate.net/profile/Irene-Coin/publication/5768317_Solid-phase_peptide_synthesis_from_standard_procedures_to_the_synthesis_of_difficult_sequences/links/00b49522da37a6a10b000000/Solid-phase-peptide-synthesis-from-standard-procedures-to-the-synthesis-of-difficult-sequences.pdf
- https://www.nature.com/articles/s41467-018-06798-7