Results: 3

1.
Figure 3

Figure 3. Neutralization of ovTNFα by seven different alpaca anti-ovTNFα VHHs. From: Alpaca (Lama pacos) as a convenient source of recombinant camelid heavy chain antibodies (VHHs).

The assay was performed as described in Material and Methods. Negative (non-immune) and positive (ovTNFα) alpaca serum was used at 1:100 dilution. Each ovTNFα VHHs was tested at 1 ug/ml concentration. G7, F4 and B5 (closed symbols) showed ovTNFα neutralizing activity while C4, C3, D3 and F1 (open symbols) had no significant neutralizing activity. Data were verified in two additional independent experiments.

David R. Maass, et al. J Immunol Methods. ;324(1-2):13-25.
2.
Figure 1

Figure 1. Characterization of alpaca serum immunoglobulins. From: Alpaca (Lama pacos) as a convenient source of recombinant camelid heavy chain antibodies (VHHs).

A. Coomassie blue stained gel following SDS-PAGE of serum from two different normal alpacas under reducing conditions. MW markers are shown (M) and their sizes indicated. B. Western blot of the alpaca serum resolved in A and probed with anti-llama IgG (H+L). The identities of the stained proteins are indicated. C and D. Coomassie blue stained gel following SDS-PAGE of three purified alpaca IgG isotypes resolved under nonreducing (Panel C; 7.5% gel) and reducing conditions (Panel D; 10% gel).

David R. Maass, et al. J Immunol Methods. ;324(1-2):13-25.
3.
Figure 2

Figure 2. Amino terminal coding sequence of VHH cDNAs and PCR primer design. From: Alpaca (Lama pacos) as a convenient source of recombinant camelid heavy chain antibodies (VHHs).

A. The cDNA encoding the complete amino terminus of 24 random VHH proteins obtained by 5’RACE from normal alpacas. The sequence of the leader and the beginning of the first framework domain are shown. The asterisks below the alignment indicate the positions of 100% nucleotide conservation. Two PCR primers (AlpVh-L and AlpVh-F1) are shown that were designed for amplification of VHH coding DNA based on the alpaca cDNA sequences. PCR primer VH1BACK, which has often been used for PCR amplification of camel and llama VHH coding DNA, is also shown. B. Agarose gel of the PCR amplification products of 23 diverse alpaca VHH cDNAs. PCRs were performed together, each with a single unique cDNA clone as the template. Each reaction contained a mixture of two CH2 ‘reverse’ primers (AlpVHH-R1 and AlpVHH-R2) with either AlpVh-F1 or VH1BACK as the ‘forward’ primer.

David R. Maass, et al. J Immunol Methods. ;324(1-2):13-25.

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