Custom Antibodies

With partner facilities based in North America where the local climate is mild all year round allowing animals to be housed in “open air” facilities, our antibody services are ideal for good quality, robust products with satisfaction guaranteed. This is because the natural environment, along with constant attention to the health and comfort of the animals, leads to a better immune response and higher quality antibody production.

The advantages of using our services:

• Custom antibodies produced at licensed facilities in the North America
• Short protocols available for research antibody development
• Longer custom antibody protocols available for large quantity needs
• Antigen design including analysis of peptide sequence, hydrophobicity, secondary structure, and antigenicity
• Fast, high quality peptide synthesis also performed within the United States
• Certified animal technicians with over 20+ years of experience
• All custom antibody development services are confidential
• Newly constructed, state-of-the-art ascites production facility

Monoclonal Antibody Production Service

• Hybridoma Generation - using automated high throughput automated platform that allows for more efficient clone testing at a very affordable price. This approach benefits from far less redundancy than the normal limit dilution method, ie, creating more diversified clones. Email us for more information
• Recombinant Rabbit Monclonals - Our partner labs use molecular engineering  to provide rabbit monoclonal antibodies without the use of a fusion partner. Thus, a very practical way to benefit from the rabbit’s immune system. Please email us for more information at Info@2BScientific.com 

Polyclonal Antibody Production Service

• Rabbit - with on-site veterinarians to continual monitor animal health & comfort, our facility is able to offer standard 60 day rabbit production packages which consist of 4 immunizations, and 3 production bleeds, typically using 2 New Zealand White Rabbits for each project. Protocols can be adjusted to suit specific needs. Email us for more information at Info@2BScientific.com
• Goat - standard 98 day protocols, with 5 immunisations and 3 bleeds per goat per project. Email us for more information at Info@2BScientific.com
• Chicken - Standard 60 protocol at the conclusion of which  0.5 gram purified (>90% pure) IgY are provided.  along with a pre-immune IgY sample. This is designed to provide a quick supply of sufficient custom chicken antibody to carry out research project. An immunisation/production schedule is provided for every project.
• See our selection of Anti-Chicken Immunoglobulin Secondary Reagents. Hens are housed and eggs are collected for an additional 2 weeks following delivery. At this time, the customer may choose to purchase more IgY. Please email us for more information at Info@2BScientific.com 

Hybridoma Sequencing and Development

quick and professional hybridoma sequencing service, also known as antibody sequencing. Hybridoma sequencing is the sequencing of the cDNA encoding the variable heavy (VH) and variable light (VL) domains of your hybridoma cell line. Email us for more information at Info@2BScientific.com

Antibody Purification

whether its monoclonal or polyclonal, a wide selection of sample types can be purified from ascites, sera to tissue culture supernatant ranging from a [For m few mg to multi-gram quantities. Email us for more information at Info@2BScientific.com

• Ammonium sulphate precipitation
• Protein A chromatography
• Protein G chromatography
• Antigen affinity purification
• PEG Purification

Antibody Conjugation

The ability to conjugate most antibodies with:

• HRP
• Biotin
• Fluorescent Dyes (FITC, Cy3, Cy5, Alexa Fluor etc.

Custom IgY

1.  For clients interested in making antibodies against highly conserved gene products (where the sequence of the human homolog is almost identical with other mammals, including rabbit)), using a host animal as far away evolutionarily from mammals is best — they recognize even the most highly conserved mammalian gene products as foreign, and have correspondingly vigorous immune reactions (birds are 300 million years removed from mammals).  I guess you could also make antibodies in sharks, but they’re not as cooperative as chickens are when you try to bleed them.

2.  We purify antibodies from eggs, rather than serum.  This means that we never have to bleed, restrain or handle the hens (except during the injections, which they don’t seem to mind, and when we need to euthanize them at the end of the project).  In contrast, bleeding rabbits involves restraints and needles poked into them (including their hearts at the end of the project).

3.  It allows folks to easily perform multiplex assays, such as double- or triple-staining.  The secondaries against chicken IgY don’t cross react at all with goat or rabbit secondary reagents (e.g., fluorescein labeled goat anti-chicken IgY).  It also lowers the cost of these secondary reagents, since their production doesn’t require pre-absorbing against other antibodies (e.g., goat anti-chicken IgY’s don’t need to be absorbed against rabbit, mouse, and human IgG’s).

4.  They are stable for long periods of time at room temperature.  This is, of course, how we can pass them out during the SfN meetings — we don’t need a booth refrigerator and recipients don’t need a hotel refrigerator.  This comes as a great surprise to most researchers, who are used to refrigeration requirements.  We simply tell them to keep the vials out of the sun (bright lights DO affect all antibodies), and put them in their refrigerator in the lab, when they get home.

5.  They cost less because collection of the raw material requires MUCH less technician time.  A technician can collect the eggs from 1000 hens in the same time it takes to bleed a rabbit.

6.  Binding properties are about the same as with rabbit IgG’s — same equilibrium Kd values (a measure of binding tightness), and kinetic on- and off-rates.  They’re no better, but no worse, too.

But, a couple of caveats:  Although chicken antibodies can be used in all of the same applications as rabbit polyclonal IgG’s, there are a couple of things clients need to know.

1.  For westerns, they need to stay away from PVDF membranes, which seem to bind chicken antibodies non-specifically.  Nitrocellulose works great, but it is a little more brittle than PVDF (but costs less).

2.  For paraformaldehyde-fixed paraffin embedded sections in IHC, it should be noted that chicken antibodies are larger than rabbit or mouse IgG’s, so penetration into tissues becomes more of an issue.  We recommend that, if their normal protocols don’t work, they should consider (a) lowering the paraformaldehyde fixation concentration to 3%, instead of 4%; and (b) include an “antigen retrieval” step before addition of primary antibody.  For example, treatment of paraffin-embedded sections with 90-95 degree heat for 20 minutes (in sodium citrate buffer) usually does the trick.

Email us for more information at Info@2BScientific.com

Contract Production Of Antibodies

Bio X Cell provides production and purification services of antibodies produced from pre-existing hybridoma cell lines. These hybridomas are typically developed in the client’s laboratory or available in the public domain.

This service includes fermentation of the hybridoma in cell culture, harvest of the cell culture supernatant by centrifugation and concentration by 30 kDa ultrafiltration. Purification is performed by Protein A or G affinity chromatography. The purified antibody is dialyzed into PBS and filter sterilized. All of the custom products will be delivered with an endotoxin level < 2EU/mg. They also offer freezing of the cell line, on-site cell line storage and MAP testing for an additional fee.

All hybridomas submitted for contract production will first be evaluated in a small-scale pilot to determine whether the productivity is suitable to proceed to full scale. The fee for the pilot is $800 but will be waived if the production proceeds to full scale.

Email us for more information at Info@2BScientific.com