Absolutely; you are welcome to visit us at any time during standard business hours (9 am to 6 pm).
We are happy to take your project order over the phone, by email, or via our website. To start the project, we require a Purchase Order or Signed Quote. For customer convenience, the majority of our services have published pricing online, which many clients find useful to generate a Purchase Order.
For California-based clients, we do charge your local sales tax rate for projects that have physical deliverables (ex. DNA, protein, cell line).
Yes. Please provide this number to us prior to the completion of your project.
We are always happy to put in place any necessary agreements before starting any service projects. We can start with one of our template documents, such as our standard Master Service Agreement or Non-Disclosure Agreement; however, we can also work from a template that you provide to us.
We are able to start most projects immediately after receipt of a Purchase Order and all required materials.
In general, we prefer a Purchase Order; however, we charge a 3.5% service fee if the client still prefers to pay with a credit card.
Most DNA cloning projects are completed in 1-2 weeks; custom projects may take longer.
For most projects, 1-2 μg of template/vector DNA is sufficient.
The preferred method of sending plasmid DNA to LakePharma is to send it in deionized water (dH20).
For all standard cloning projects (PCR cloning, ligation cloning, site mutagenesis, Gateway cloning) the deliverable is miniprep DNA and a Certificate of Analysis. Projects that include sequence confirmation will also receive a sequence confirmation file showing an alignment to your gene of interest. Endotoxin-free DNA scale-up (midiprep, maxiprep) is available upon request for an additional cost.
While we would prefer that we receive sufficient quantities of DNA to perform all cloning steps, if needed we can transform and perform a miniprep of the vector.
We can make AAV, Lentivirus, and baculovirus.
There are multiple ways to submit the sample:
- Freeze the antibody producing cells and send us a frozen vial on dry ice (1 million cells is ideal, though 100,000 is sufficient).
- Send frozen purified total RNA from the antibody producing cells.
- Send the total RNA sample from the antibody producing cells that has been dried on an RNA isolation column and not eluted.
Please fill out a Sample Submission Form for each sample, including the IgG isotype if known.
Our standard hybridoma sequencing procedure gives us the full variable region and part of the constant region of heavy and light chains. We provide an antibody sequence analysis report specifying the number of heavy and light chains identified, analysis of the results, agarose gel images of the amplified DNA fragments, and the DNA and protein sequences for each chain identified. Construction of vectors to express the antibody genes and binding confirmation are additional services that can be performed following hybridoma sequencing.
If you do not know which light or heavy chain is correct by looking at the sequence, we recommend expressing all possible combinations of the heavy and light chains in small-scale, followed by assaying all antibodies for activity (either from the conditioned media or from a purified sample).
Including DNA vector construction, most small-scale production projects are completed in 3 weeks.
Yes, we actually prefer if you let us know ahead of time the number and size of each vial, and whether you require a specific formulation buffer.
Yes, the concentration can be adjusted depending on your needs. Please note that some proteins may precipitate when they are concentrated too much, which could drastically reduce the final yield.
Why some proteins express better than others is complex; protein expression can be influenced by factors beyond transcription and translation. At LakePharma we have many tools to optimize sequences. Autologous mRNA optimization encompasses aspects beyond eliminating rare codons, including increasing GC content, eliminating hairpins, repeats, cryptic splice sites, and premature polyadenylation signals, all of which could make the correct message more abundant, stable, and amenable to translation. Performing this type of mRNA optimization may result in higher expression of some transcripts/proteins; however, there is no guarantee that this optimization will work in all cases.
We offer a full suite of custom protein purification options, including affinity chromatography (ex. antibody, immobilized-metal, and others), ion exchange chromatography, hydrophobic interaction chromatography, size exclusion chromatography, and more.
Yes. Any native protein purification would be a custom project; please contact us to discuss your needs.
Protein concentration is determined by A280 Nanodrop, adjusting for the theoretical extinction coefficient.
CHO stable cell lines expressing an antibody or protein become economical for many reasons:
- The antibody/protein is a very low expresser, since our stable cell line process can increase the titer of some proteins by 5-10 fold or more
- You require a large amount of antibody/protein (several hundred mg to gram scale)
- You want to produce the same antibody multiple times at large quantities over time
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