Previously, we highlighted the impressive capabilities of the Ella automated ELISA system. While Ella offers speed and automation, it's important to recognize the enduring value of traditional ELISA (enzyme-linked immunosorbent assay) methods as well. Let's take a closer look at why ELISA remains a trusted and effective option in many research settings. Despite the rise of automation, there are situations where precision, flexibility, and cost-efficiency make ELISA the preferred choice.
Advantages of ELISA
Cost Considerations: ELISA methods generally require a lower initial equipment investment. For laboratories with limited budgets or those handling smaller amounts of samples, ELISA is a practical and economical choice.
Customizability and Flexibility: ELISA allows for greater flexibility in assay design and customization. Researchers who need to develop and optimize their own assays, particularly for novel or less common targets, can benefit from ELISA development as it allows for more control over each step of the process.
Limited Sample Throughput: In situations where there are only a few samples that need to be processed, the automation and high-throughput capabilities of the Ella system may not be necessary, making ELISA a more practical choice.
Specialized Research Needs: Certain specialized research applications may require modifications or specific conditions that are easier to manage or implement with ELISA. For example, using unique reagents or specific incubation conditions that are not compatible with the automated system of Ella.
While Ella offers a high degree of automation and efficiency, ELISA methods still hold their ground in scenarios where cost and customization are the driving factors.
Example of Unique ELISA development
SBH was approached to develop an ELISA for a novel biomarker that is not commercially available on the Ella, Luminex, or ELISA kits. SBH found a few different antibodies for this biomarker as well as recombinant proteins that are commercially available. SBH developed the ELISA by initially screening all the antibodies in a direct ELISA, where the recombinant protein is bound to the plate and the antibodies bind to the recombinant protein. Additional analysis of the antibodies confirmed which antibody could be utilized as a capture antibody.
Graph 1
Three different antibodies were tested as detection antibodies (labeled as Detection Ab. A-C in Graph 1). Detection Antibody A had the most signal with the capture antibody and specific recombinant protein, as seen above.
Next, SBH had to confirm that the signal observed is specific to the target of interest. The target of interest shares 98% similarity to another analyte. There was concern that the ELISA would detect this analyte in addition to the novel target, so SBH confirmed the specificity of the recombinant protein and detection antibody. First, SBH tested both the recombinant protein of the novel target as well as the recombinant protein of the similar analyte to confirm the detection antibody only recognized the novel biomarker of interest.
Graph 2
Detection antibody A only recognized the novel target recombinant protein and not the other analyte (Graph 2).
Graph 3
Additionally, an antibody for the other analyte (Detection Antibody D) was tested against the recombinant protein of interest to confirm that it is only recognized by the specific antibody for this target. (Graph 3) SBH confirmed that any signal observed is specific in this manner.
The next step in the ELISA development was confirming if the expected sample type could be tested in this ELISA. The samples to be tested with this novel biomarker were cell lysates. SBH tested a few different lysis buffers and a few different concentrations of the lysis buffers.
Graph 4
The result was that the 50% lysis buffer masked the signal, while the 25% lysis buffer was similar to the diluent. (Graph 4)
This is an example of how SBH can provide custom ELISA solutions to meet your specific needs.
SBH Sciences: Your Trusted ELISA Partner
At SBH Sciences, we can perform any type of ELISA you require, including assays that accurately measure the concentration of antigens or antibodies. Whether you need direct, indirect, sandwich, or competitive ELISA, our team is equipped to deliver precise, high-quality results.
We have successfully developed over 30 highly sensitive and accurate colorimetric sandwich ELISA kits for the measurement of cytokines and biomarkers for research and development purposes, as well as to assess PK/PD during clinical trials. Examples of our work include kits for cytokines (TNF-α, VEGF, IL-12 p70), biomarkers/cancer (Galectin-1, Galectin-3), and apoptosis markers (Bcl-1, Caspase-3). Furthermore, we offer in-house optimization to examine the impacts of different factors on your PK/PD studies. We are currently optimizing ELISA conditions to enhance drastically ELISA sensitivity.
We would be delighted to assist you.
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